Differential knockout of a heterozygous allele of lrrk2

ABSTRACT

RNA molecules comprising a guide sequence portion having 17-50 contiguous nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and compositions, methods, and uses thereof.

This application claims the benefit of U.S. Provisional Application Nos. 63/213,506, filed Jun. 22, 2021, and 63/119,905, filed Dec. 1, 2020, the contents of which are hereby incorporated by reference.

Throughout this application, various publications are referenced, including referenced in parenthesis. The disclosures of all publications mentioned in this application in their entireties are hereby incorporated by reference into this application in order to provide additional description of the art to which this invention pertains and of the features in the art which can be employed with this invention.

REFERENCE TO SEQUENCE LISTING

This application incorporates-by-reference nucleotide sequences which are present in the file named “211201_91648-A-PCT_Sequence_Listing_AWG.txt”, which is 19,634 kilobytes in size, and which was created on Nov. 29, 2021 in the IBM-PC machine format, having an operating system compatibility with MS-Windows, which is contained in the text file filed Dec. 1, 2021 as part of this application.

BACKGROUND OF INVENTION

There are several classes of DNA variation in the human genome, including insertions and deletions, differences in the copy number of repeated sequences, and single nucleotide polymorphisms (SNPs). A SNP is a DNA sequence variation occurring when a single nucleotide (adenine (A), thymine (T), cytosine (C), or guanine (G)) in the genome differs between human subjects or paired chromosomes in an individual. Over the years, the different types of DNA variations have been the focus of the research community either as markers in studies to pinpoint traits or disease causation or as potential causes of genetic disorders.

A genetic disorder is caused by one or more abnormalities in the genome. Genetic disorders may be regarded as either “dominant” or “recessive.” Recessive genetic disorders are those which require two copies (i.e., two alleles) of the abnormal/defective gene to be present. In contrast, a dominant genetic disorder involves a gene or genes which exhibit(s) dominance over a normal (functional/healthy) gene or genes. As such, in dominant genetic disorders only a single copy (i.e., allele) of an abnormal gene is required to cause or contribute to the symptoms of a particular genetic disorder. Such mutations include, for example, gain-of-function mutations in which the altered gene product possesses a new molecular function or a new pattern of gene expression. Other examples include dominant negative mutations, which have a gene product that acts antagonistically to the wild-type allele.

Leucine-Rich Repeat Serine/Threonine-Protein Kinase 2 (LRRK2) and Monogenic Parkinson's Disease (PD)

Leucine-rich repeat serine/threonine-protein kinase 2 (LRRK2) is a generalist kinase that phosphorylates a broad range of proteins involved in multiple processes, such as neuronal plasticity, autophagy, and vesicle trafficking. It is a key regulator of Rab GTPases such as the Rab-3 family (Rab-3A, Rab-3B, Rab-3C, Rab-3D), the Rab-5 family (Rab-5A, Rab-5B, Rab-5C), the Rab-8 family (Rab-8A, Rab-8B), Rab-10, Rab-12, Rab-35, and Rab-43. It also regulates their interaction partners by extension in modulating the exchange between GDP and GTP on these proteins. These interacting partners govern a wide variety of cellular processes, including autophagy, cell differentiation, cell transport, synaptic vesicle trafficking, retrograde trafficking pathways for protein recycling, neuronal morphology in the central nervous system (CNS), and inhibiting degradation of the microtubule-associated protein tau (MAPT).

The LRRK2 G2019S mutation (p.Gly2019Ser, c.6055G>A) is the most common PD-associated mutation known. Indeed, LRRK2 G2019S was reported in 41% of sporadic and 37% of familial PD patients (compared to 3% in healthy controls) from the North African Arab population, and in 18.3% of Ashkenazi Jewish PD patients (compared to 1.3% in controls).

While the precise pathogenic mechanism is unknown, the characterized mutations (such as Gly2019Ser in exon 41) appear to disrupt kinase or GTPase activity. These disruptions in activity seem to result in increased intra- and intermolecular phosphorylation of known LRRK2 substrates. LRRK2-mediated PD is similar to idiopathic PD, which is a disease characterized by progressive degeneration of motor functions, progressive asymmetric tremors at rest, muscle rigidity, postural instability, REM sleep behavior disorders, cognitive decline, and gait abnormalities. Some of these symptoms are less severe in LRRK2-mediated PD patients than they are for idiopathic PD patients. Onset for LRRK2-mediated PD is typically after age 50, though early-onset (20s) and late-onset (90s) versions of the disease have been described. The LRRK2 gene consists of 51 exons, with the coding region starting on exon 1, and the G1y2019Ser mutation occurs in exon 41, in what is known as the “activation hinge” of the kinase domain.

SUMMARY OF THE INVENTION

Disclosed is an approach for knocking out the expression of a dominant-mutated LRRK2 allele by disrupting the dominant-mutated allele or degrading the resulting mRNA.

The present disclosure provides a method for utilizing at least one naturally occurring nucleotide difference or polymorphism (e.g., single nucleotide polymorphism (SNP)) for distinguishing/discriminating between two alleles of a gene, one allele bearing a mutation such that it encodes a mutated protein causing a disease phenotype (“mutated allele”) and a particular sequence in a SNP position (REF/SNP), and the other allele encoding for a functional protein (“functional allele”). In some embodiments, the SNP position is utilized for distinguishing/discriminating between two alleles of a gene bearing one or more disease associated mutations, such as to target one of the alleles bearing both the particular sequence in the SNP position (SNP/REF) and a disease associated mutation. In some embodiments, the disease-associated mutation is targeted. In some embodiments, the method further comprises the step of knocking out expression of the mutated protein and allowing expression of the functional protein.

The present disclosure also provides a method for modifying in a cell a mutant allele of the Leucine-rich repeat serine/threonine-protein kinase 2 (LRRK2) gene having a mutation associated with monogenic Parkinson's disease, the method comprising

-   -   introducing to the cell a composition comprising:         -   a CRISPR nuclease or a sequence encoding the CRISPR             nuclease; and         -   a first RNA molecule comprising a guide sequence portion             having 17-50 nucleotides or a nucleotide sequence encoding             the same,     -   wherein a complex of the CRISPR nuclease and the first RNA         molecule affects a double strand break in the mutant allele of         the LRRK2 gene.

According to embodiments of the present invention, there is provided a first RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID Nos: 1-103,606.

According to some embodiments of the present invention, there is provided a composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease.

According to some embodiments of the present invention, there is provided a method for inactivating a mutant LRRK2 allele in a cell, the method comprising delivering to the cell a composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease. In some embodiments, the cell is a stem cell. In some embodiments, the cell is a neuron. In some embodiments, the cell is a dopaminergic cell. In some embodiments, the delivering to the cell is performed in vitro, ex vivo, or in vivo. In some embodiments, the method is performed ex vivo and the cell is provided/explanted from an individual patient. In some embodiments, the method further comprises the step of introducing the resulting cell, with the modified/knocked out mutant LRRK2 allele, into the individual patient (e.g., autologous transplantation).

According to some embodiments of the present invention, there is provided a method for treating monogenic Parkinson's disease, the method comprising delivering to a cell of a subject having monogenic Parkinson's disease a composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease.

According to some embodiments of the present invention, there is provided use of a composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease for inactivating a mutant LRRK2 allele in a cell, comprising delivering to the cell the composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease.

According to embodiments of the present invention, there is provided a medicament comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease for use in inactivating a mutant LRRK2 allele in a cell, wherein the medicament is administered by delivering to the cell the composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease.

According to some embodiments of the present invention, there is provided use of a composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease for treating ameliorating or preventing monogenic Parkinson's disease, comprising delivering to a cell of a subject having or at risk of having monogenic Parkinson's disease the composition of comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease. In some embodiments, the method is performed ex vivo and the cell is provided/explanted from the subject. In some embodiments, the method further comprises the step of introducing the resulting cell, with the modified/knocked out mutant LRRK2 allele, into the subject (e.g. autologous transplantation).

According to some embodiments of the present invention, there is provided a medicament comprising the composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease for use in treating ameliorating or preventing monogenic Parkinson's disease, wherein the medicament is administered by delivering to a cell of a subject having or at risk of having monogenic Parkinson's disease the composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease.

According to some embodiments of the present invention, there is provided a kit for inactivating a mutant LRRK2 allele in a cell, comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606, a CRISPR nuclease, and/or a tracrRNA molecule; and instructions for delivering the RNA molecule; CRISPR nuclease, and/or the tracrRNA to the cell.

According to some embodiments of the present invention, there is provided a kit for treating monogenic Parkinson's disease in a subject, comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606, a CRISPR nuclease, and/or a tracrRNA molecule; and instructions for delivering the RNA molecule; CRISPR nuclease, and/or the tracrRNA to a cell of a subject having or at risk of having monogenic Parkinson's disease.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 : Editing activity of guides targeting LRRK2 in A549 cells. RNPs complexed with SpCas9 CRISPR nuclease and a guide molecule comprising a specific guide sequence portion were electroporated into A549 cells to determine their activity. Cells were harvested 72 hours post RNP electroporation, genomic was DNA extracted, and the region of the mutation was amplified and analyzed by next-generation sequencing (NGS). The graph represents the % of editing±standard deviation of two (2) independent electroporations. The accompanying table below the graph indicates the genotype of A549 cells at each SNP position.

FIG. 2 : Relative amount of LRRK2 RNA. A549 cells were harvested seven (7) days post-electroporation, total RNA was extracted, and the extracted RNA was subjected to reverse transcription. The level of the generated cDNA was quantified using qRT-PCR relative to untreated cells that were not edited and normalized to an endogenous gene (GAPDH). The graph represents the % of editing±standard deviation of two (2) independent electroporations.

FIG. 3 : Editing activity of guides targeting LRRK2 in HeLa cells with OMNI-159.

FIG. 4 : Editing activity of guides targeting LRRK2 in HeLa cells with OMNI-79 V5570.

DETAILED DESCRIPTION

Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

It should be understood that the terms “a” and “an” as used above and elsewhere herein refer to “one or more” of the enumerated components. It will be clear to one of ordinary skill in the art that the use of the singular includes the plural unless specifically stated otherwise. Therefore, the terms “a,” “an” and “at least one” are used interchangeably in this application.

For purposes of better understanding the present teachings and in no way limiting the scope of the teachings, unless otherwise indicated, all numbers expressing quantities, percentages or proportions, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

Unless otherwise stated, adjectives such as “substantially” and “about” modifying a condition or relationship characteristic of a feature or features of an embodiment of the invention, are understood to mean that the condition or characteristic is defined to within tolerances that are acceptable for operation of the embodiment for an application for which it is intended. Unless otherwise indicated, the word “or” in the specification and claims is considered to be the inclusive “or” rather than the exclusive or, and indicates at least one of, or any combination of items it conjoins.

In the description and claims of the present application, each of the verbs, “comprise,” “include” and “have” and conjugates thereof, are used to indicate that the object or objects of the verb are not necessarily a complete listing of components, elements or parts of the subject or subjects of the verb. Other terms as used herein are meant to be defined by their well-known meanings in the art.

The terms “nucleic acid template” and “donor”, refer to a nucleotide sequence that is inserted or copied into a genome. The nucleic acid template comprises a nucleotide sequence, e.g., of one or more nucleotides, that will be added to or will template a change in the target nucleic acid or may be used to modify the target sequence. A nucleic acid template sequence may be of any length, for example between 2 and 10,000 nucleotides in length, preferably between about 100 and 1,000 nucleotides in length, more preferably between about 200 and 500 nucleotides in length. A nucleic acid template may be a single stranded nucleic acid, a double stranded nucleic acid. In some embodiments, the nucleic acid template comprises a nucleotide sequence, e.g., of one or more nucleotides, that corresponds to wild type sequence of the target nucleic acid, e.g., of the target position. In some embodiments, the nucleic acid template comprises a nucleotide sequence, e.g., of one or more ribonucleotides, that corresponds to wild type sequence of the target nucleic acid, e.g., of the target position. In some embodiments, the nucleic acid template comprises modified nucleotides.

Insertion of an exogenous sequence (also called a “donor sequence,” donor template,” “donor molecule” or “donor”) can also be carried out. For example, a donor sequence can contain a non-homologous sequence flanked by two regions of homology to allow for efficient HDR at the location of interest. Additionally, donor sequences can comprise a vector molecule containing sequences that are not homologous to the region of interest in cellular chromatin. A donor molecule can contain several, discontinuous regions of homology to cellular chromatin. For example, for targeted insertion of sequences not normally present in a region of interest, said sequences can be present in a donor nucleic acid molecule and flanked by regions of homology to sequence in the region of interest. A donor molecule may be any length, for example ranging from several bases e.g. 10-20 bases to multiple kilobases in length.

The donor polynucleotide can be DNA or RNA, single-stranded and/or double-stranded and can be introduced into a cell in linear or circular form. See, e.g., U.S. Patent Publication Nos. 2010/0047805; 2011/0281361; 2011/0207221; and 2019/0330620. If introduced in linear form, the ends of the donor sequence can be protected (e.g., from exonucleolytic degradation) by methods known to those of skill in the art. For example, one or more dideoxynucleotide residues are added to the 3′ terminus of a linear molecule and/or self-complementary oligonucleotides are ligated to one or both ends. See, for example, Chang et al. (1987) and Nehls et al. (1996). Additional methods for protecting exogenous polynucleotides from degradation include, but are not limited to, addition of terminal amino group(s) and the use of modified intemucleotide linkages such as, for example, phosphorothioates, phosphoramidates, and O-methyl ribose or deoxyribose residues.

A donor sequence may be an oligonucleotide and be used for targeted alteration of an endogenous sequence. The oligonucleotide may be introduced to the cell on a vector, may be electroporated into the cell, or may be introduced via other methods known in the art. Donor polynucleotides can be introduced as naked nucleic acid, as nucleic acid complexed with an agent such as a liposome or poloxamer, or can be delivered by viruses (e.g., adenovirus, AAV, herpesvirus, retrovirus, lentivirus and integrase defective lentivirus (IDLY)).

As used herein, the term “modified cells” refers to cells in which a double strand break is affected by a complex of an RNA molecule and the CRISPR nuclease as a result of hybridization with the target sequence, i.e. on-target hybridization. The term “modified cells” may further encompass cells in which a repair or correction of a mutation was affected following the double strand break.

This invention provides a modified cell or cells obtained by use of any of the methods described herein. In an embodiment these modified cell or cells are capable of giving rise to progeny cells. In an embodiment these modified cell or cells are capable of giving rise to progeny cells after engraftment. As a non-limiting example, the modified cells may be hematopoietic stem cells (HSCs), or any cell suitable for an allogenic cell transplant or autologous cell transplant. As a non-limiting example, the modified cells may be stem cells, neurons, or dopaminergic cells.

This invention also provides a composition comprising these modified cells and a pharmaceutically acceptable carrier. Also provided is an in vitro or ex vivo method of preparing this, comprising mixing the cells with the pharmaceutically acceptable carrier.

As used herein, the term “targeting sequence” or “targeting molecule” refers a nucleotide sequence or molecule comprising a nucleotide sequence that is capable of hybridizing to a specific target sequence, e.g., the targeting sequence has a nucleotide sequence which is at least partially complementary to the sequence being targeted along the length of the targeting sequence. The targeting sequence or targeting molecule may be part of an RNA molecule that can form a complex with a CRISPR nuclease, either alone or in combination with other RNA molecules, with the targeting sequence serving as the targeting portion of the CRISPR complex. When the molecule having the targeting sequence is present contemporaneously with the CRISPR nuclease, the RNA molecule, alone or in combination with an additional one or more RNA molecules (e.g. a tracrRNA molecule), is capable of targeting the CRISPR nuclease to the specific target sequence. As non-limiting example, a guide sequence portion of a CRISPR RNA molecule or single-guide RNA molecule may serve as a targeting molecule. Each possibility represents a separate embodiment. A targeting sequence can be custom designed to target any desired sequence.

The term “targets” as used herein, refers to preferentially hybridizing a targeting sequence of a targeting molecule to a nucleic acid having a targeted nucleotide sequence. It is understood that the term “targets” encompasses variable hybridization efficiencies, such that there is preferential targeting of the nucleic acid having the targeted nucleotide sequence, but unintentional off-target hybridization in addition to on-target hybridization might also occur. It is understood that where an RNA molecule targets a sequence, a complex of the RNA molecule and a CRISPR nuclease molecule targets the sequence for nuclease activity.

The “guide sequence portion” of an RNA molecule refers to a nucleotide sequence that is capable of hybridizing to a specific target DNA sequence, e.g., the guide sequence portion has a nucleotide sequence which is partially or fully complementary to the DNA sequence being targeted along the length of the guide sequence portion. In some embodiments, the guide sequence portion is 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 nucleotides in length, or approximately 17-50, 17-49, 17-48, 17-47, 17-46, 17-45, 17-44, 17-43, 17-42, 17-41, 17-40, 17-39, 17-38, 17-37, 17-36, 17-35, 17-34, 17-33, 17-31, 17-30, 17-29, 17-28, 17-27, 17-26, 17-25, 17-24, 17-22, 17-21, 18-25, 18-24, 18-23, 18-22, 18-21, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-22, 18-20, 20-21, 21-22, or 17-20 nucleotides in length. Preferably, the entire length of the guide sequence portion is fully complementary to the DNA sequence being targeted along the length of the guide sequence portion. The guide sequence portion may be part of an RNA molecule that can form a complex with a CRISPR nuclease with the guide sequence portion serving as the DNA targeting portion of the CRISPR complex. When the RNA molecule having the guide sequence portion is present contemporaneously with the CRISPR molecule, alone or in combination with an additional one or more RNA molecules (e.g. a tracrRNA molecule), the RNA molecule is capable of targeting the CRISPR nuclease to the specific target DNA sequence. Accordingly, a CRISPR complex can be formed by direct binding of the RNA molecule having the guide sequence portion to a CRISPR nuclease or by binding of the RNA molecule having the guide sequence portion and an additional one or more RNA molecules to the CRISPR nuclease. Each possibility represents a separate embodiment. A guide sequence portion can be custom designed to target any desired sequence. Accordingly, a molecule comprising a “guide sequence portion” is a type of targeting molecule. In some embodiments, the guide sequence portion comprises a sequence that is the same as, or differs by no more than 1, 2, 3, 4, or 5 nucleotides from, a guide sequence portion described herein, e.g., a guide sequence set forth in any of SEQ ID NOs:1-103,606. Each possibility represents a separate embodiment. In some embodiments, the guide sequence portion comprises a sequence that is the same as, or differs by up to 1, 2, 3, 4, or 5 nucleotides from, a guide sequence portion described herein, e.g., a guide sequence set forth in any of SEQ ID NOs:1-103,606. Each possibility represents a separate embodiment. In some of these embodiments, the guide sequence portion comprises a sequence that is the same as a sequence set forth in any of SEQ ID NOs:1-103,606. Throughout this application, the terms “guide molecule,” “RNA guide molecule,” “guide RNA molecule,” and “gRNA molecule” are synonymous with a molecule comprising a guide sequence portion.”

The term “non-discriminatory” as used herein refers to a guide sequence portion of an RNA molecule that targets a specific DNA sequence that is common both a mutant and functional allele of a gene.

In embodiments of the present invention, an RNA molecule comprises a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606.

The RNA molecule and or the guide sequence portion of the RNA molecule may contain modified nucleotides. Exemplary modifications to nucleotides or polynucleotides may be synthetic and encompass polynucleotides which contain nucleotides comprising bases other than the naturally occurring adenine, cytosine, thymine, uracil, or guanine bases. Modifications to polynucleotides include polynucleotides which contain synthetic, non-naturally occurring nucleosides e.g., locked nucleic acids. Modifications to polynucleotides may be utilized to increase or decrease stability of an RNA. An example of a modified polynucleotide is an mRNA containing 1-methyl pseudo-uridine. For examples of modified polynucleotides and their uses, see U.S. Pat. No. 8,278,036, PCT International Publication No. WO/2015/006747, and Weissman and Kariko (2015), each of which is hereby incorporated by reference.

As used herein, “contiguous nucleotides” set forth in a SEQ ID NO refers to nucleotides in a sequence of nucleotides in the order set forth in the SEQ ID NO without any intervening nucleotides.

In embodiments of the present invention, the guide sequence portion may be 25 nucleotides in length and contain 20-22 contiguous nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606. In embodiments of the present invention, the guide sequence portion may be less than 22 nucleotides in length. For example, in embodiments of the present invention the guide sequence portion may be 17, 18, 19, 20, or 21 nucleotides in length. In such embodiments the guide sequence portion may consist of 17, 18, 19, 20, or 21 nucleotides, respectively, in the sequence of 17-22 contiguous nucleotides set forth in any one of SEQ ID NOs: 1-103,606. For example, a guide sequence portion having 17 nucleotides in the sequence of 17 contiguous nucleotides set forth in SEQ ID NO: 103,605 may consist of any one of the following nucleotide sequences (nucleotides excluded from the contiguous sequence are marked in strike-through):

(SEQ ID NO: 103, 607) AAAAAAAUGUACUUGGUUCC 17 nucleotide guide sequence 1: (SEQ ID NO: 103, 608)

AAAAUGUACUUGGUUCC 17 nucleotide guide sequence 2: (SEQ ID NO: 103, 609)

AAAAAUGUACUUGGUUC

17 nucleotide guide sequence 3: (SEQ ID NO: 103, 610)

AAAAAAUGUACUUGGUU

17 nucleotide guide sequence 4: (SEQ ID NO: 103, 611) AAAAAAAUGUACUUGGU

In embodiments of the present invention, the guide sequence portion may be greater than 20 nucleotides in length. For example, in embodiments of the present invention the guide sequence portion may be 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In such embodiments the guide sequence portion comprises 17-50 nucleotides containing the sequence of 20, 21 or 22 contiguous nucleotides set forth in any one of SEQ ID NOs: 1-103,606 and additional nucleotides fully complimentary to a nucleotide or sequence of nucleotides adjacent to the 3′ end of the target sequence, 5′ end of the target sequence, or both.

In embodiments of the present invention a CRISPR nuclease and an RNA molecule comprising a guide sequence portion form a CRISPR complex that binds to a target DNA sequence to effect cleavage of the target DNA sequence. CRISPR nucleases, e.g. Cpf1, may form a CRISPR complex comprising a CRISPR nuclease and RNA molecule without a further tracrRNA molecule. Alternatively, CRISPR nucleases, e.g. Cas9, may form a CRISPR complex between the CRISPR nuclease, an RNA molecule, and a tracrRNA molecule. A guide sequence portion, which comprises a nucleotide sequence that is capable of hybridizing to a specific target DNA sequence, and a sequence portion that participates in CRIPSR nuclease binding, e.g. a tracrRNA sequence portion, can be located on the same RNA molecule. Alternatively, a guide sequence portion may be located on one RNA molecule and a sequence portion that participates in CRIPSR nuclease binding, e.g. a tracrRNA portion, may located on a separate RNA molecule. A single RNA molecule comprising a guide sequence portion (e.g. a DNA-targeting RNA sequence) and at least one CRISPR protein-binding RNA sequence portion (e.g. a tracrRNA sequence portion), can form a complex with a CRISPR nuclease and serve as the DNA-targeting molecule. In some embodiments, a first RNA molecule comprising a DNA-targeting RNA portion, which includes a guide sequence portion, and a second RNA molecule comprising a CRISPR protein-binding RNA sequence interact by base pairing to form an RNA complex that targets the CRISPR nuclease to a DNA target site or, alternatively, are fused together to form an RNA molecule that complexes with the CRISPR nuclease and targets the CRISPR nuclease to a DNA target site.

In embodiments of the present invention, a RNA molecule comprising a guide sequence portion may further comprise the sequence of a tracrRNA molecule. Such embodiments may be designed as a synthetic fusion of the guide portion of the RNA molecule and the trans-activating crRNA (tracrRNA). (See Jinek et al., 2012). In such an embodiment, the RNA molecule is a single guide RNA (sgRNA) molecule. Embodiments of the present invention may also form CRISPR complexes utilizing a separate tracrRNA molecule and a separate RNA molecule comprising a guide sequence portion. In such embodiments the tracrRNA molecule may hybridize with the RNA molecule via basepairing and may be advantageous in certain applications of the invention described herein.

The term “tracr mate sequence” refers to a sequence sufficiently complementary to a tracrRNA molecule so as to hybridize to the tracrRNA via basepairing and promote the formation of a CRISPR complex. (See U.S. Pat. No. 8,906,616). In embodiments of the present invention, the RNA molecule may further comprise a portion having a tracr mate sequence.

A “gene,” for the purposes of the present disclosure, includes a DNA region encoding a gene product, as well as all DNA regions which regulate the production of the gene product, whether or not such regulatory sequences are adjacent to coding and/or transcribed sequences. Accordingly, a gene includes, but is not necessarily limited to, promoter sequences, terminators, translational regulatory sequences such as ribosome binding sites and internal ribosome entry sites, enhancers, silencers, insulators, boundary elements, replication origins, matrix attachment sites and locus control regions.

“Eukaryotic” cells include, but are not limited to, fungal cells (such as yeast), plant cells, animal cells, mammalian cells and human cells.

The term “nuclease” as used herein refers to an enzyme capable of cleaving the phosphodiester bonds between the nucleotide subunits of nucleic acid. A nuclease may be isolated or derived from a natural source. The natural source may be any living organism. Alternatively, a nuclease may be a modified or a synthetic protein which retains the phosphodiester bond cleaving activity. Gene modification can be achieved using a nuclease, for example a CRISPR nuclease.

As used herein, the term “HSC” refers to both hematopoietic stem cells and hematopoietic stem progenitor cells. Non-limiting examples of stem cells include bone marrow cells, myeloid progenitor cells, a multipotent progenitor cells, and lineage restricted progenitor cells.

As used herein, “progenitor cell” refers to a lineage cell that is derived from stem cell and retains mitotic capacity and multipotency (e.g., can differentiate or develop into more than one but not all types of mature lineage of cell). As used herein “hematopoiesis” or “hemopoiesis” refers to the formation and development of various types of blood cells (e.g., red blood cells, megakaryocytes, myeloid cells (e.g., monocytes, macrophages and neutrophil), and lymphocytes) and other formed elements in the body (e.g., in the bone marrow).

The term “single nucleotide polymorphism (SNP) position”, as used herein, refers to a position in which a single nucleotide DNA sequence variation occurs between members of a species, or between paired chromosomes in an individual. In the case that a SNP position exists at paired chromosomes in an individual, a SNP on one of the chromosomes is a “heterozygous SNP.” The term SNP position refers to the particular nucleic acid position where a specific variation occurs and encompasses both a sequence including the variation from the most frequently occurring base at the particular nucleic acid position (also referred to as “SNP” or alternative “ALT”) and a sequence including the most frequently occurring base at the particular nucleic acid position (also referred to as reference, or “REF”). Accordingly, the sequence of a SNP position may reflect a SNP (i.e. an alternative sequence variant relative to a consensus reference sequence within a population), or the reference sequence itself.

According to embodiments of the present invention, there is provided a method for modifying in a cell a mutant allele of the Leucine-rich repeat serine/threonine-protein kinase 2 (LRRK2) gene having a mutation associated with monogenic Parkinson's disease, the method comprising

-   -   introducing to the cell a composition comprising:         -   at least one CRISPR nuclease or a sequence encoding a CRISPR             nuclease; and         -   a first RNA molecule comprising a guide sequence portion             having 17-50 nucleotides or a nucleotide sequence encoding             the same,     -   wherein a complex of the CRISPR nuclease and the first RNA         molecule affects a double strand break in the mutant allele of         the LRRK2 gene.

In some embodiments, the first RNA molecule targets the CRISPR nuclease to the mutation associated with monogenic Parkinson's disease.

In some embodiments, the mutation associated with monogenic Parkinson's disease is any one of 12:40225041_C_T, 12:40225058_C_A, 12:40225088_G_A, 12:40225159_G_A, 12:40225233_G_C, 12:40225558_C_T, 12:40225618_T_C, 12:40225628_G_A, 12:40225631_T_C, 12:40232303_A_C, 12:40232345_G_C, 12:40232380_A_C, 12:40232385_T_C, 12:40235634_T_C, 12:40235646_C_G, 12:40235660_A_G, 12:40238078_A_G, 12:40240517_C_T, 12:40240531_T_C, 12:40240594_G_C, 12:40240602_T_C, 12:40243551_CAAT_C, 12:40243556_A_T, 12:40249822_T_G, 12:40249843_C_G, 12:40249854_C_T, 12:40249881_T_C, 12:40249908_A_G, 12:40251273_G_A, 12:40251303_G_A, 12:40251361_A_G, 12:40251366_C_A, 12:40251369_G_A, 12:40251495_C_T, 12:40252906_A_G, 12:40252929_G_A, 12:40252939_C_T, 12:40252952_T_G, 12:40252958_A_C, 12:40252968_G_A, 12:40252974_C_T, 12:40252978_C_T, 12:40252984_C_T, 12:40252988_T_C, 12:40253016_G_A, 12:40257311_A_G, 12:40257341_G_T, 12:40259507_A_G, 12:40259525_A_T, 12:40259534_G_T, 12:40259578_G_A, 12:40263806_A_G, 12:40263875_A_G, 12:40263900_G_A, 12:40263907_T_C, 12:40274646_CC_C, 12:40274881_T_C, 12:40274899_A_G, 12:40274917_G_C, 12:40277933_T_C, 12:40277968_A_C, 12:40277971_A_G, 12:40278025_T_A, 12:40278154_A_G, 12:40278167_C_T, 12:40283881_G_A, 12:40283915_T_C, 12:40283947_C_T, 12:40283966_G_A, 12:40283985_C_A, 12:40283990_T_C, 12:40284061_A_G, 12:40284076_C_A, 12:40284139_T_C, 12:40287363_C_A, 12:40287438_A_G, 12:40287461_A_G, 12:40293555_CTC_CCT, 12:40293607_C_T, 12:40293613_G_A, 12:40293624_G_C, 12:40293626_G_A, 12:40293644_A_G, 12:40294854_T_G, 12:40294862_T_C, 12:40294866_G_T, 12:40294878_C_T, 12:40295448_A_G, 12:40295463_A_G, 12:40295466_G_A, 12:40295535_A_T, 12:40295566_A_G, 12:40295574_A_C, 12:40295581_G_T, 12:40298330_A_G, 12:40298333_C_T, 12:40298346_G_A, 12:40298433_C_G, 12:40298479_G_T, 12:40298488_A_G, 12:40299125_A_G, 12:40299190_C_T, 12:40299196_A_C, 12:40299212_G_A, 12:40299255_T_C, 12:40302866_A_G, 12:40303952_C_A, 12:40303972_C_G, 12:40304000_G_A, 12:40304004_A_G, 12:40304035_A_C, 12:40304040_G_C, 12:40304040_G_T, 12:40304078_G_A, 12:40304141_C_T, 12:40305776_C_T, 12:40305781_T_A, 12:40305791_C_G, 12:40305871_G_A, 12:40305942_G_A, 12:40305946_T_A, 12:40308467_G_T, 12:40308476_A_G, 12:40308481_G_A, 12:40308543_GG_G, 12:40308632_C_A, 12:40308672_G_A, 12:40309174_G_A, 12:40309206_C_T, 12:40309225_A_C, 12:40309226_A_G, 12:40309230_A_G, 12:40309236_G_A, 12:40310434_C_A, 12:40310434_C_G, 12:40310434_C_T, 12:40310435_G_A, 12:40310436_C_T, 12:40310437_G_C, 12:40310440_T_G, 12:40310450_C_T, 12:40310461_G_A, 12:40310471_A_G, 12:40310476_GAT_G, 12:40310515_A_G, 12:40310530_C_T, 12:40310546_G_A, 12:40310554_G_A, 12:40310561_G_A, 12:40314043_G_A, 12:40314059_C_T, 12:40314062_A_G, 12:40314101_C_A, 12:40315266_T_A, 12:40319979_A_T, 12:40319998_T_C, 12:40320043_G_A, 12:40320054_G_C, 12:40320090_A_G, 12:40320094_A_G, 12:40320097_T_C, 12:40320119_A_G, 12:40321114_A_G, 12:40321123_T_C, 12:40322037_C_T, 12:40322038_G_A, 12:40322047_G_A, 12:40322047_G_T, 12:40322134_A_G, 12:40322137_A_C, 12:40322364_T_C, 12:40322386_G_T, 12:40322468_C_A, 12:40323244_C_G, 12:40323255_A_G, 12:40323256_T_C, 12:40323260_G_T, 12:40323270_G_T, 12:40323297_T_C, 12:40328416_G_C, 12:40335008_A_T, 12:40335031_G_A, 12:40335036_C_T, 12:40335037_G_A, 12:40335079_G_T, 12:40340349_C_G, 12:40340361_T_C, 12:40340380_T_C, 12:40340400_G_A, 12:40340404_T_C, 12:40340432_A_G, 12:40340436_A_T, 12:40346818_T_C, 12:40346826_ACTACT_A, 12:40346884_A_G, 12:40348402_G_GTGT, 12:40348475_A_G, 12:40348484_C_T, 12:40351537_A_G, 12:40351572_T_A, 12:40351579_C_T, 12:40351585_G_A, 12:40351605_G_C, 12:40351623_G_T, 12:40351667_C_A, 12:40351680_G_C, 12:40351720_G_C, 12:40351723_A_G, 12:40354304_T_C, 12:40354305_G_T, 12:40354410_G_A, 12:40356126_A_T, 12:40356145_C_T, 12:40359345_C_T, 12:40359422_A_G, 12:40363435_C_T, 12:40363440_C_T, 12:40363541_G_A, 12:40364843_G_A, 12:40364845_G_GGT, 12:40364884_G_A, 12:40364927_A_T, 12:40364960_A_G, 12:40365044_C_T, 12:40367012_T_A, 12:40367050_A_G, 12:40367648_AC_A, 12:40367656_G_T, 12:40367754_T_C, 12:40367849_G_A, 12:40367888_A_C, 12:40367988_C_T, 12:40368042_A_G, 12:40368073_A_C, 12:40368170_C_T, 12:40368262_C_T, 12:40368380_T_A, 12:40368486_G_T, 12:40368516_C_A, 12:40368534_T_G, 12:40368673_A_C, 12:40368736_C_A, 12:40368783_G_T, 12:40368800_T_C, 12:40368807_A_G, 12:40368834_A_G, 12:40368860_C_T, 12:40368905_A_T, 12:40368930_C_T, 12:40368947_C_T, 12:40368951_C_T, 12:40369044_T_C, 12:40369153_T_C, and 12:40369167_G_T.

In some embodiments, the guide sequence portion of the first RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 that targets a mutation associated with monogenic Parkinson's disease.

In some embodiments, the first RNA molecule targets the CRISPR nuclease to a SNP position of the mutant allele.

In some embodiments, the SNP position is any one of rs1388598, rs28365207, rs7294619, rs10878226, rs1491943, rs1491942, rs1491941, rs11175645, rs4567538, rs12230685, rs11175655, rs11175656, rs1402299410, rs780970090, rs1479481931, rs71078229, rs11175658, rs3057613, rs35642658, rs74324737, rs2708441, rs10631840, rs35991926, rs61931634, rs11175666, rs751672176, rs36024911, rs377568728, rs1279923177, rs2249281, rs10878244, rs138000936, rs10878245, rs10878246, rs10878247, rs954884, rs10878249, rs10784444, rs6581622, rs17519573, rs35987733, rs868476079, rs1399756142, rs1474763022, rs1450276224, rs1191097078, rs1425272990, rs1477924701, rs2404580, rs2404581, rs1388599, rs1388596, rs10878258, rs12371502, rs11175729, rs17443414, rs1486599371, rs7134379, rs7955902, rs1491939, rs1491938, rs7969677, rs2723265, rs7132171, rs10784451, rs10784452, rs1907633, rs1491936, rs1907632, rs2046928, rs2723264, rs17465870, rs10878278, rs11175766, rs11564183, rs11175769, rs17465912, rs17443552, rs17490817, rs11430638, rs4293189, rs10784461, rs7308720, rs10784462, rs10878284, rs11175780, rs11175784, rs11175787, rs144463374, rs73108329, rs12426891, rs4768224, rs11175793, rs10784470, rs12146857, rs11564207, rs7308193, rs10506150, rs76660489, rs10878299, rs373619252, rs10878302, rs11564129, rs7971935, rs11175836, rs4272849, rs10784485, rs10506151, rs7309197, rs7134572, rs11175843, rs147801391, rs376096297, rs7488873, rs11175845, rs7979341, rs1226327150, rs777452337, rs1339001348, rs11175847, rs112859457, rs17466080, rs11175852, rs10784486, rs73108368, rs398019201, rs17519846, rs17466150, rs11175862, rs4310676, rs11564150, rs4318033, rs4473003, rs1429478461, rs886049356, rs11564270, rs17519881, rs17466178, rs17466185, rs11175902, rs76033741, rs56325408, rs7957754, rs11175911, rs12369882, rs11175913, rs4483664, rs66487867, rs1491426750, rs5006481, rs139565575, rs150276517, rs72446556, rs1186032972, rs12820920, rs7966550, rs12314692, rs11175922, rs73102746, rs10784497, rs17443815, rs7957057, rs76607433, rs28365224, rs7962011, rs10878336, rs11175934, rs11175935, rs17491061, rs11175941, rs10878340, rs10784498, rs10878341, rs10784499, rs17484286, rs146071397, rs10878343, rs11564180, rs11175958, rs11175959, rs113772607, rs11564149, rs11175963, rs7305344, rs7133914, rs11175964, rs397805328, rs11175966, rs10784501, rs41286474, rs869212703, rs2069228, rs10784503, rs386376250, rs11564128, rs2896975, rs7302503, rs7137160, rs369794148, rs34709132, rs10467144, rs11175985, rs10784504, rs57421387, rs7973058, rs1427267, rs1427266, rs721713, rs721712, rs721711, rs721709, rs4768227, rs398098159, rs4767969, rs10878356, rs10784506, rs10878358, rs7132073, rs1427265, rs17466339, rs2114569, rs2114568, rs2162469, rs10784509, rs1896254, rs5797667, rs373676201, rs1896253, rs1896252, rs1427263, rs11176013, rs11564148, rs11564205, rs10748030, rs10748031, rs10748032, rs11564177, rs10748033, rs10735931, rs10748034, rs10878368, rs7307276, rs4768229, rs4768230, rs397968969, rs11564204, rs4423250, rs7137665, rs10878371, rs11176022, rs60750674, rs10878372, rs398055492, rs761931834, rs1415940047, rs17466430, rs11176030, rs3747891, rs7298930, rs17444089, rs10878377, rs12316801, rs17444096, rs11564175, rs2404832, rs17466486, rs11564267, rs17520244, rs10506152, rs11176052, rs11176053, rs17444124, rs11564203, r57307562, rs17520251, rs11461395, rs10506153, rs73275761, rs963243, rs17444145, rs10878386, rs17466521, rs12367542, rs17520278, rs35031086, rs11834950, rs11829088, rs4465428, rs61007767, rs60776971, rs2404834, rs7302841, rs2404835, rs11356084, rs715403, rs715402, rs6581667, rs6581668, rs200521371, rs4767970, rs4767971, rs4767972, rs1427271, rs1427272, rs1427273, rs10735934, rs10732751, rs12306060, rs1365763, rs7963697, rs11316380, rs7963086, rs17491536, rs7956787, rs7956898, rs10506155, rs7954061, rs7957151, rs919174, rs3065921, rs919175, rs1035812, rs1365764, rs397724721, rs17461964, rs10784518, rs4768231, rs4768232, rs3943893, rs2162471, rs1035813, rs10459264, rs10459265, rs11564147, rs10715759, rs7296657, rs7312497, rs10715758, rs10784522, rs796827140, rs74467833, rs7131694, rs12311273, rs58911468, rs35847030, rs1365765, rs66513412, rs2896977, rs199606989, rs1365766, rs10878405, rs11176143, rs7294958, rs61915610, rs7294952, rs59096461, rs59980086, rs4768233, rs4768234, rs4768235, rs7314455, rs7313525, rs7313895, rs7314863, rs7303525, rs7132187, rs7307310, rs11176153, rs11176154, rs7311247, rs7137437, rs75439407, rs73107108, rs1307806388, rs10878409, rs10878410, rs138382676, rs11176161, rs10878412, rs10878413, rs183544696, rs189538849, rs11610569, rs142726158, rs1194726090, rs576444406, rs185723137, rs1179215763, rs71449759, rs61915613, rs140722234, rs191099092, rs113693842, rs112767992, rs34531686, rs12368227, rs12368229, rs3930031, rs7308626, rs890575, rs890575, rs10748040, rs61230125, rs11176165, rs7976715, rs7976724, rs7963987, rs2162472, rs767501990, rs552326490, rs1365381164, rs760911090, rs754028820, rs11564173, rs7306545, rs7306684, rs112259589, rs73277531, rs77689380, rs73277533, rs17462055, rs758880647, rs10784528, rs796949072, rs10784532, rs145580704, rs386376254, rs11836288, rs11564146, rs10784536, rs11277474, rs7971919, rs7962116, rs67512688, rs2896978, rs2404836, rs11176195, rs4767973, rs17444202, rs4768236, rs4768237, rs58724345, rs397897325, rs33962975, rs7134408, rs11835105, rs137858615, rs10878434, rs35403247, rs3761863, rs17444223, rs12426362, rs17466605, rs3789328, rs12426498, rs4768238, rs66737902, rs10878441, rs3886747, rs1365770, rs367772598, rs386376255, rs10784548, rs1465527, rs1465528, rs17444285, rs7968048, rs11609433, rs776238897, rs149833872, rs1299545494, rs4374003, and rs7962370.

In some embodiments, the guide sequence portion of the first RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 that targets a SNP position of the mutant allele.

In some embodiments, the SNP position is in an exon or intron of the LRRK2 mutant allele, or within 3,000 basepairs upstream of the transcription start site or within 3,000 basepairs downstream of the 3′ UTR of the LRRK2 mutant allele.

In some embodiments, the SNP position contains a heterozygous SNP.

In some embodiments, the method further comprises introducing to the cell a second RNA molecule comprising a guide sequence portion having 17-50 nucleotides or a nucleotide sequence encoding the same, wherein a complex of the second RNA molecule and a CRISPR nuclease affects a second double strand break in the LRRK2 gene.

In some embodiments, the guide sequence portion of the second RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 other than the sequence of the first RNA molecule.

In some embodiments, the second RNA molecule comprises a non-discriminatory guide portion that targets both functional and mutated LRRK2 alleles.

In some embodiments, the second RNA molecule comprises a non-discriminatory guide portion that targets any one of Intron 1 of LRRK2, Intron 49 of LRRK2, Intron 2 of LRRK2, Intron 3 of LRRK2, a 3′ untranslated region (3′ UTR) of LRRK2, and an intergenic region downstream of LRRK2.

In some embodiments, the second RNA molecule comprises a non-discriminatory guide portion that targets a sequence that is located within a genomic range selected from any one of 12:40224997-40225496, 12:40225283-40225554, 12:40366506-40367005, 12:40367078-40367643, 12:40225641-40226140, and 12:40235126-40235625.

In some embodiments, the second RNA molecule comprises a non-discriminatory guide portion that targets a sequence that is located up to 500 base pairs from an exon that is excised by the first and second RNA molecules.

In some embodiments, an exon or a portion thereof is excised from the mutant allele of the LRRK2 gene.

In some embodiments, the first RNA molecule targets a SNP position in the 3′ UTR of the mutated allele, and the second RNA molecule comprises a non-discriminatory guide portion that targets downstream of a polyadenylation signal sequence that is common to both a functional allele and the mutant allele of the LRRK2 gene.

In some embodiments, the first RNA molecule targets a SNP position downstream of a polyadenylation signal of the mutated allele, and the second RNA molecule comprises a non-discriminatory guide portion that targets a sequence upstream of a polyadenylation signal that is common to both a functional allele and the mutant allele of the LRRK2 gene.

In some embodiments, the polyadenylation signal is excised from the mutant allele of the LRRK2 gene.

In some embodiments, the guide sequence portion of the first RNA molecule or second RNA molecule comprises 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion.

In some embodiments, the guide sequence portion comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion.

In some embodiments, the guide sequence portion provides higher targeting specificity to the complex of the CRISPR nuclease and the first RNA molecule relative to a guide sequence portion that has higher complementarity to the mutant allele of the LRRK2 gene

According to embodiments of the present invention, there is provided a modified cell obtained by the method of any one of the embodiments presented herein. In some embodiments the modified cell is a stem cell, a neuron, or a dopaminergic cell.

According to embodiments of the present invention, there is provided a first RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606, or any one of SEQ ID NOs: 1-103,606 modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion.

According to embodiments of the present invention, there is provided a composition comprising the first RNA molecule and at least one CRISPR nuclease.

In some embodiments, the composition further comprises a second RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides, wherein the second RNA molecule targets a LRRK2 allele, and wherein the guide sequence portion of the second RNA molecule is a different sequence from the sequence of the guide sequence portion of the first RNA molecule.

In some embodiments, the guide sequence portion of the second RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 other than the sequence of the first RNA molecule, or any one of SEQ ID NOs: 1-103,606 modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion.

According to embodiments of the present invention, there is provided a method for inactivating a mutant LRRK2 allele in a cell, the method comprising delivering to the cell the composition of any one of the embodiments presented herein.

According to embodiments of the present invention, there is provided a method for treating monogenic Parkinson's disease, the method comprising delivering to a cell of a subject having monogenic Parkinson's disease the composition of any one of the embodiments presented herein.

According to embodiments of the present invention, there is provided use of any one of the compositions presented herein for inactivating a mutant LRRK2 allele in a cell, comprising delivering to the cell the composition of any one of the embodiments presented herein.

According to embodiments of the present invention, there is provided a medicament comprising the composition of any one of the embodiments presented herein for use in inactivating a mutant LRRK2 allele in a cell, wherein the medicament is administered by delivering to the cell the composition of any one of the embodiments presented herein.

According to embodiments of the present invention, there is provided use of the composition of any one of the embodiments presented herein for treating ameliorating or preventing monogenic Parkinson's disease, comprising delivering to a cell of a subject having or at risk of having monogenic Parkinson's disease the composition of any one of the embodiments presented herein.

According to embodiments of the present invention, there is provided a medicament comprising the composition of any one of the embodiments presented herein for use in treating ameliorating or preventing monogenic Parkinson's disease, wherein the medicament is administered by delivering to a cell of a subject having or at risk of having monogenic Parkinson's disease the composition of any one of the embodiments presented herein.

According to embodiments of the present invention, there is provided a kit for inactivating a mutant LRRK2 allele in a cell, comprising an RNA molecule of any one of the embodiments presented herein, a CRISPR nuclease, and/or a tracrRNA molecule; and instructions for delivering the RNA molecule; CRISPR nuclease, and/or the tracrRNA to the cell.

According to embodiments of the present invention, there is provided a kit for treating monogenic Parkinson's disease in a subject, comprising an RNA molecule of any one of the embodiments presented herein, a CRISPR nuclease, and/or a tracrRNA molecule; and instructions for delivering the RNA molecule; CRISPR nuclease, and/or the tracrRNA to a cell of a subject having or at risk of having monogenic Parkinson's disease.

According to embodiments of the present invention, there is provided a gene editing composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606. In some embodiments, the RNA molecule further comprises a portion having a sequence which binds to a CRISPR nuclease. In some embodiments, the sequence which binds to a CRISPR nuclease is a tracrRNA sequence.

In some embodiments, the RNA molecule further comprises a portion having a tracr mate sequence.

In some embodiments, the RNA molecule may further comprise one or more linker portions.

According to embodiments of the present invention, an RNA molecule may be up to 1000, 900, 800, 700, 600, 500, 450, 400, 350, 300, 290, 280, 270, 260, 250, 240, 230, 220, 210, 200, 190, 180, 170, 160, 150, 140, 130, 120, 110, or 100 nucleotides in length. Each possibility represents a separate embodiment. In embodiments of the present invention, the RNA molecule may be 17 up to 300 nucleotides in length, 100 up to 300 nucleotides in length, 150 up to 300 nucleotides in length, 100 up to 500 nucleotides in length, 100 up to 400 nucleotides in length, 200 up to 300 nucleotides in length, 100 to 200 nucleotides in length, or 150 up to 250 nucleotides in length. Each possibility represents a separate embodiment.

According to some embodiments of the present invention, the composition further comprises a tracrRNA molecule.

The present disclosure provides a method for utilizing at least one naturally occurring nucleotide difference or polymorphism (e.g., single nucleotide polymorphism (SNP)) for distinguishing/discriminating between two alleles of a gene, one allele bearing a mutation such that it encodes a mutated protein causing a disease phenotype (“mutated allele”) and a particular sequence in a SNP position (SNP/REF), and the other allele encoding for a functional protein (“functional allele”). The method further comprises the step of knocking out expression of the mutated protein and allowing expression of the functional protein. In some embodiments, the method is for treating, ameliorating, or preventing a dominant negative genetic disorder.

According to some embodiments of the present invention, there is provided a method for inactivating a mutant LRRK2 allele in a cell, the method comprising delivering to the cell a composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease.

According to some embodiments of the present invention, there is provided a method for treating monogenic Parkinson's disease, the method comprising delivering to a cell of a subject having monogenic Parkinson's disease a composition comprising an RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 and a CRISPR nuclease.

According to embodiments of the present invention, the composition comprises a second RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606. In some embodiments, the 17-50 nucleotides of the guide sequence portion of the second RNA molecule are in a different sequence from the sequence of the guide sequence portion of the first RNA molecule.

According to embodiments of the present invention, at least one CRISPR nuclease and the RNA molecule or RNA molecules are delivered to the subject and/or cells substantially at the same time or at different times.

In some embodiments, a tracrRNA molecule is delivered to the subject and/or cells substantially at the same time or at different times as the CRISPR nuclease and RNA molecule or RNA molecules.

According to embodiments of the present invention, the first RNA molecule targets a SNP or disease-causing mutation in the exon or promoter of a mutated allele, and the second RNA molecule targets a SNP in an exon of the mutated allele, a SNP in an intron, or a sequence present in both the mutated or functional allele.

According to embodiments of the present invention, the first RNA molecule or the first and the second RNA molecules target a SNP in the promoter region, the start codon, or an untranslated region (UTR) of a mutated allele.

According to embodiments of the present invention, the first RNA molecule or the first and the second RNA molecules targets at least a portion of the promoter and/or the start codon and/or a portion of a UTR of a mutated allele.

According to embodiments of the present invention, the first RNA molecule targets a portion of the promoter, a first SNP in the promoter, or a SNP upstream to the promoter of a mutated allele and the second RNA molecule is targets a second SNP, which is downstream of the first SNP, and is in the promoter, in a UTR, or in an intron or in an exon of a mutated allele.

According to embodiments of the present invention, the first RNA molecule targets a SNP in the promoter, upstream of the promoter, or a UTR of a mutated allele and the second RNA molecule is designed to target a sequence which is present in an intron of both the mutated allele and the functional allele.

According to embodiments of the present invention, the first RNA molecule targets a SNP in an intron of a mutated allele, and wherein the second RNA molecule targets a SNP in an intron of the mutated allele, or a sequence in an intron present in both the mutated and functional allele.

According to embodiments of the present invention, the first RNA molecule targets a sequence upstream of the promotor which is present in both a mutated and functional allele and the second RNA molecule targets a SNP or disease-causing mutation in any location of the gene.

According to embodiments of the present invention, there is provided a method comprising removing an exon containing a disease-causing mutation from a mutated allele, wherein the first RNA molecule or the first and the second RNA molecules target regions flanking an entire exon or a portion of the exon.

According to embodiments of the present invention, there is provided a method comprising removing an exon or a portion thereof from a mutant LRRK2 allele, the entire open reading frame of a mutant LRRK2 allele, or removing the entire mutant LRRK2 allele.

According to embodiments of the present invention, the first RNA molecule targets a SNP or disease-causing mutation in an exon or promoter of a mutated allele, and wherein the second RNA molecule targets a SNP in the same exon of the mutated allele, a SNP in an intron, or a sequence in an intron present in both the mutated and functional allele.

According to embodiments of the present invention, the first RNA molecule or the first and the second RNA molecules target an alternative splicing signal sequence between an exon and an intron of a mutant LRRK2 allele.

According to embodiments of the present invention, the second RNA molecule is non-discriminatory targets a sequence present in both a mutated allele and a functional allele.

The compositions and methods of the present disclosure may be utilized for treating, preventing, ameliorating, or slowing progression of an autosomal dominant genetic disorder, such as monogenic Parkinson's disease.

In some embodiments, a mutated allele is deactivated by delivering to a cell an RNA molecule which targets a SNP in the promoter region, the start codon, or an untranslated region (UTR) of the mutated allele.

In some embodiments, a mutated allele is inactivated by removing at least a portion of the promoter, and/or removing the start codon, and/or a portion of the UTR, and/or a polyadenylation signal. In such embodiments one RNA molecule may be designed for targeting a first SNP in the promoter or upstream to the promoter and another RNA molecule is designed to target a second SNP, which is downstream of the first SNP, and is in the promoter, in the UTR, in an intron, or in an exon. Alternatively, one RNA molecule may be designed for targeting a SNP in the promoter, upstream of the promoter, or the UTR, and another RNA molecule is designed to target a sequence which is present in an intron of both the mutated allele and the functional allele. Alternatively, one RNA molecule may be designed for targeting a sequence upstream of the promotor which is present in both the mutated and functional allele and the other guide is designed to target a SNP or disease-causing mutation in any location of the gene e.g., in an exon, intron, UTR, or downstream of the promoter.

In some embodiments, the method of deactivating a mutated allele comprises an exon skipping step comprising removing an exon containing a disease-causing mutation from the mutated allele. Removing an exon containing a disease-causing mutation in the mutated allele requires two RNA molecules which target regions flanking the entire exon or a portion of the exon. Removal of an exon containing the disease-causing mutation may be designed to eliminate the disease-causing action of the protein while allowing for expression of the remaining protein product which retains some or all of the wild-type activity. The entire open reading frame or the entire gene can be excised using two RNA molecules flanking the region desired to be excised.

In some embodiments, the method of deactivating a mutated allele comprises delivering two RNA molecules to a cell, wherein one RNA molecule targets a SNP or disease-causing mutation in an exon or promoter of the mutated allele, and wherein the other RNA molecule targets a SNP in the same of the mutated allele, a SNP in an intron, or a sequence in an intron present in both the mutated or functional allele.

Any one of, or combination of, the above-mentioned strategies for deactivating a mutant allele may be used in the context of the invention.

In embodiments of the present invention, an RNA molecule is used to direct a CRISPR nuclease to an exon or a splice site of a mutated allele in order to create a double-stranded break (DSB), leading to insertion or deletion of nucleotides by inducing an error-prone non-homologous end-joining (NHEJ) mechanism and formation of a frameshift mutation in the mutated allele. The frameshift mutation may result in, for example, inactivation or knockout of the mutated allele by generation of an early stop codon in the mutated allele and to generation of a truncated protein or to nonsense-mediated mRNA decay of the transcript of the mutant allele. In further embodiments, one RNA molecule is used to direct a CRISPR nuclease to a promotor of a mutated allele.

In some embodiments, the method of deactivating a mutated allele further comprises enhancing activity of the functional protein such as by providing a protein/peptide, a nucleic acid encoding a protein/peptide, or a small molecule such as a chemical compound, capable of activating/enhancing activity of the functional protein.

According to some embodiments, the present disclosure provides an RNA sequence (also referred to as an ‘RNA molecule’) which binds to or associates with and/or directs an RNA-guided DNA nuclease e.g., a CRISPR nuclease, to a target sequence comprising at least one nucleotide which differs between a mutated allele and a functional allele (e.g., SNP) of a gene of interest (i.e., a sequence of the mutated allele which is not present in the functional allele).

In some embodiments, the method comprises contacting a mutated allele of a gene of interest with an allele-specific RNA molecule and a CRISPR nuclease e.g., a Cas9 protein, wherein the allele-specific RNA molecule and the CRISPR nuclease associate with a nucleotide sequence of the mutated allele of the gene of interest which differs by at least one nucleotide from a nucleotide sequence of a functional allele of the gene of interest, thereby modifying or knocking-out the mutated allele.

In some embodiments, the allele-specific RNA molecule and a CRISPR nuclease is introduced to a cell encoding the gene of interest. In some embodiments, the cell encoding the gene of interest is in a mammalian subject. In some embodiments, the cell encoding the gene of interest is in a plant.

In some embodiments, the mutated allele is an allele of LRRK2 gene. In some embodiments, the RNA molecule targets a SNP which co-exists with or is genetically linked to the mutated sequence associated with monogenic Parkinson's disease genetic disorder. In some embodiments, the RNA molecule targets a SNP which is highly prevalent in the population and exists in the mutated allele having the mutated sequence associated with monogenic Parkinson's disease genetic disorder and not in the functional allele of an individual subject to be treated. In some embodiments, a disease-causing mutation within a mutated LRRK2 allele is targeted.

In some embodiments, the SNP is within an exon of the gene of interest. In such embodiments, a guide sequence portion of an RNA molecule is designed to associate with a sequence of an exon of the gene of interest.

In some embodiments, SNP is within an intron or the exon of the gene of interest. In some embodiments, the SNP is in close proximity to the splice site between an intron and an exon. In some embodiments, the close proximity to a splice site is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides upstream or downstream to the splice site. Each possibility represents a separate embodiment of the present invention. In such embodiments, a guide sequence portion of an RNA molecule may be designed to associate with a sequence of the gene of interest which comprises the splice site.

In some embodiments, the method is utilized for treating a subject having a disease phenotype resulting from the heterozygote LRRK2 gene. In such embodiments, the method results in improvement, amelioration or prevention of the disease phenotype.

Embodiments of compositions described herein include at least one CRISPR nuclease, RNA molecule(s), and a tracrRNA molecule, being effective in a subject or cells at the same time. The at least one CRISPR nuclease, RNA molecule(s), and tracrRNA may be delivered substantially at the same time or can be delivered at different times but have effect at the same time. For example, this includes delivering the CRISPR nuclease to the subject or cells before the RNA molecule and/or tracrRNA is substantially extant in the subject or cells.

In some embodiments, the cell is a stem cell. In some embodiments, the cell is a neuron. In some embodiments, the cell is a dopaminergic cell.

Dominant Genetic Disorders

One of skill in the art will appreciate that all subjects with any type of heterozygote genetic disorder (e.g., dominant genetic disorder) may be subjected to the methods described herein. In one embodiment, the present invention may be used to target a gene involved in, associated with, or causative of dominant genetic disorders such as, for example, monogenic Parkinson's disease. In some embodiments, the dominant genetic disorder is monogenic Parkinson's disease. In some embodiments, the target gene is the LRRK2 gene. Non-limiting examples of mutations characterized as gain of function mutations associated with monogenic Parkinson's disease phenotype include: 12:40225041_C_T, 12:40225058_C_A, 12:40225088_G_A, 12:40225159_G_A, 12:40225233_G_C, 12:40225558_C_T, 12:40225618_T_C, 12:40225628_G_A, 12:40225631_T_C, 12:40232303_A_C, 12:40232345_G_C, 12:40232380_A_C, 12:40232385_T_C, 12:40235634_T_C, 12:40235646_C_G, 12:40235660_A_G, 12:40238078_A_G, 12:40240517_C_T, 12:40240531_T_C, 12:40240594_G_C, 12:40240602_T_C, 12:40243551_CAAT_C, 12:40243556_A_T, 12:40249822_T_G, 12:40249843_C_G, 12:40249854_C_T, 12:40249881_T_C, 12:40249908_A_G, 12:40251273_G_A, 12:40251303_G_A, 12:40251361_A_G, 12:40251366_C_A, 12:40251369_G_A, 12:40251495_C_T, 12:40252906_A_G, 12:40252929_G_A, 12:40252939_C_T, 12:40252952_T_G, 12:40252958_A_C, 12:40252968_G_A, 12:40252974_C_T, 12:40252978_C_T, 12:40252984_C_T, 12:40252988_T_C, 12:40253016_G_A, 12:40257311_A_G, 12:40257341_G_T, 12:40259507_A_G, 12:40259525_A_T, 12:40259534_G_T, 12:40259578_G_A, 12:40263806_A_G, 12:40263875_A_G, 12:40263900_G_A, 12:40263907_T_C, 12:40274646_CC_C, 12:40274881_T_C, 12:40274899_A_G, 12:40274917_G_C, 12:40277933_T_C, 12:40277968_A_C, 12:40277971_A_G, 12:40278025_T_A, 12:40278154_A_G, 12:40278167_C_T, 12:40283881_G_A, 12:40283915_T_C, 12:40283947_C_T, 12:40283966_G_A, 12:40283985_C_A, 12:40283990_T_C, 12:40284061_A_G, 12:40284076_C_A, 12:40284139_T_C, 12:40287363_C_A, 12:40287438_A_G, 12:40287461_A_G, 12:40293555_CTC_CCT, 12:40293607_C_T, 12:40293613_G_A, 12:40293624_G_C, 12:40293626_G_A, 12:40293644_A_G, 12:40294854_T_G, 12:40294862_T_C, 12:40294866_G_T, 12:40294878_C_T, 12:40295448_A_G, 12:40295463_A_G, 12:40295466_G_A, 12:40295535_A_T, 12:40295566_A_G, 12:40295574_A_C, 12:40295581_G_T, 12:40298330_A_G, 12:40298333_C_T, 12:40298346_G_A, 12:40298433_C_G, 12:40298479_G_T, 12:40298488_A_G, 12:40299125_A_G, 12:40299190_C_T, 12:40299196_A_C, 12:40299212_G_A, 12:40299255_T_C, 12:40302866_A_G, 12:40303952_C_A, 12:40303972_C_G, 12:40304000_G_A, 12:40304004_A_G, 12:40304035_A_C, 12:40304040_G_C, 12:40304040_G_T, 12:40304078_G_A, 12:40304141_C_T, 12:40305776_C_T, 12:40305781_T_A, 12:40305791_C_G, 12:40305871_G_A, 12:40305942_G_A, 12:40305946_T_A, 12:40308467_G_T, 12:40308476_A_G, 12:40308481_G_A, 12:40308543_GG_G, 12:40308632_C_A, 12:40308672_G_A, 12:40309174_G_A, 12:40309206_C_T, 12:40309225_A_C, 12:40309226_A_G, 12:40309230_A_G, 12:40309236_G_A, 12:40310434_C_A, 12:40310434_C_G, 12:40310434_C_T, 12:40310435_G_A, 12:40310436_C_T, 12:40310437_G_C, 12:40310440_T_G, 12:40310450_C_T, 12:40310461_G_A, 12:40310471_A_G, 12:40310476_GAT_G, 12:40310515_A_G, 12:40310530_C_T, 12:40310546_G_A, 12:40310554_G_A, 12:40310561_G_A, 12:40314043_G_A, 12:40314059_C_T, 12:40314062_A_G, 12:40314101_C_A, 12:40315266_T_A, 12:40319979_A_T, 12:40319998_T_C, 12:40320043_G_A, 12:40320054_G_C, 12:40320090_A_G, 12:40320094_A_G, 12:40320097_T_C, 12:40320119_A_G, 12:40321114_A_G, 12:40321123_T_C, 12:40322037_C_T, 12:40322038_G_A, 12:40322047_G_A, 12:40322047_G_T, 12:40322134_A_G, 12:40322137_A_C, 12:40322364_T_C, 12:40322386_G_T, 12:40322468_C_A, 12:40323244_C_G, 12:40323255_A_G, 12:40323256_T_C, 12:40323260_G_T, 12:40323270_G_T, 12:40323297_T_C, 12:40328416_G_C, 12:40335008_A_T, 12:40335031_G_A, 12:40335036_C_T, 12:40335037_G_A, 12:40335079_G_T, 12:40340349_C_G, 12:40340361_T_C, 12:40340380_T_C, 12:40340400_G_A, 12:40340404_T_C, 12:40340432_A_G, 12:40340436_A_T, 12:40346818_T_C, 12:40346826_ACTACT_A, 12:40346884_A_G, 12:40348402_G_GTGT, 12:40348475_A_G, 12:40348484_C_T, 12:40351537_A_G, 12:40351572_T_A, 12:40351579_C_T, 12:40351585_G_A, 12:40351605_G_C, 12:40351623_G_T, 12:40351667_C_A, 12:40351680_G_C, 12:40351720_G_C, 12:40351723_A_G, 12:40354304_T_C, 12:40354305_G_T, 12:40354410_G_A, 12:40356126_A_T, 12:40356145_C_T, 12:40359345_C_T, 12:40359422_A_G, 12:40363435_C_T, 12:40363440_C_T, 12:40363541_G_A, 12:40364843_G_A, 12:40364845_G_GGT, 12:40364884_G_A, 12:40364927_A_T, 12:40364960_A_G, 12:40365044_C_T, 12:40367012_T_A, 12:40367050_A_G, 12:40367648_AC_A, 12:40367656_G_T, 12:40367754_T_C, 12:40367849_G_A, 12:40367888_A_C, 12:40367988_C_T, 12:40368042_A_G, 12:40368073_A_C, 12:40368170_C_T, 12:40368262_C_T, 12:40368380_T_A, 12:40368486_G_T, 12:40368516_C_A, 12:40368534_T_G, 12:40368673_A_C, 12:40368736_C_A, 12:40368783_G_T, 12:40368800_T_C, 12:40368807_A_G, 12:40368834_A_G, 12:40368860_C_T, 12:40368905_A_T, 12:40368930_C_T, 12:40368947_C_T, 12:40368951_C_T, 12:40369044_T_C, 12:40369153_T_C, and 12:40369167_G_T.

LRRK2 editing strategies include, but are not limited to, (1) knockout, for example, by targeting a LRRK2 mutation or SNP position with one guide RNA molecule to induce a frameshift that may lead to truncation or nonsense-mediated decay; and (2) allele-specific excision using two guide RNA molecules, for example, excision of at least one exon or a portion thereof, knockout of a large portion of the allele or the entire allele, or excision of the polyadenylation signal or the promoter region.

Knockout may be achieved by several approaches. For example, knockout may be achieved by targeting a SNP within a coding exon of a mutant LRRK2 allele using only one guide RNA molecule (e.g. a single guide RNA molecule or “sgRNA”). Alternatively, excision may be achieved by targeting the mutant LRRK2 allele with two different RNA molecules, with at least one RNA molecule being allele-specific.

In another editing strategy, expression of a mutated LRRK2 allele may be inhibited. An example of this strategy includes excising the polyadenylation signal in the 3′UTR region, which leads to an unstable transcript.

According to embodiments of the present invention, there is provided an RNA molecule comprising a guide sequence portion (e.g. a targeting sequence) comprising a nucleotide sequence that is fully or partially complementary to a target sequence in a SNP position (REF/SNP sequence) located in or near a mutated allele of the LRRK2 gene. In some embodiments, the guide sequence portion of the RNA molecule consists of 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more than 26 nucleotides. In some embodiments the guide sequence portion is configured to target a CRISPR nuclease to a target sequence and provide a cleavage event, by a CRISPR nuclease complexed therewith, selected from a double-strand break and a single-strand break within 500, 400, 300, 200, 100, 50, 25, or 10 nucleotides of a LRRK2 target site. In some embodiments, the cleavage event enables non-sense mediated decay of the LRRK2 gene. In some embodiments, the RNA molecule is a guide RNA molecule such as a crRNA molecule or a single guide RNA molecule.

In some embodiments, the target sequence of a mutated allele of LRRK2 gene is altered (e.g., by introduction of an NHEJ-mediated indel (e.g., insertion or deletion), and results in reduction or elimination of expression of the gene product encoded by the mutant allele of LRRK2 gene. In some embodiments, the reduction or elimination of expression is due to non-sense mediated mRNA decay such as due to immature stop codon. In some embodiments, the reduction or elimination of expression is due to expression of a truncated form of the LRRK2 gene product.

CRISPR Nucleases and PAM Recognition

In some embodiments, the sequence specific nuclease is selected from CRISPR nucleases, or a functional variant thereof. In some embodiments, the sequence specific nuclease is an RNA guided DNA nuclease. In such embodiments, the RNA sequence which guides the RNA guided DNA nuclease (e.g., Cpf1) binds to and/or directs the RNA guided DNA nuclease to the sequence comprising at least one nucleotide which differs between a mutated allele and its counterpart functional allele (e.g., SNP). In some embodiments, the CRISPR complex does not further comprise a tracrRNA. In a non-limiting example, in which the RNA guided DNA nuclease is a CRISPR protein, the at least one nucleotide which differs between the dominant mutated allele and the functional allele may be within the PAM site and/or proximal to the PAM site within the region that the RNA molecule is designed to hybridize to. A skilled artisan will appreciate that RNA molecules can be engineered to bind to a target of choice in a genome by commonly known methods in the art.

The term “PAM” as used herein refers to a nucleotide sequence of a target DNA located in proximity to the targeted DNA sequence and recognized by the CRISPR nuclease complex. The PAM sequence may differ depending on the nuclease identity. In addition, there are CRISPR nucleases that can target almost all PAMs. In some embodiments of the present invention, a CRISPR system utilizes one or more RNA molecules having a guide sequence portion to direct a CRISPR nuclease to a target DNA site via Watson-Crick base-pairing between the guide sequence portion and the protospacer on the target DNA site, which is next to the protospacer adjacent motif (PAM), which is an additional requirement for target recognition. The CRISPR nuclease then mediates cleavage of the target DNA site to create a double-stranded break within the protospacer. In a non-limiting example, a type II CRISPR system utilizes a mature crRNA:tracrRNA complex that directs the CRISPR nuclease, e.g. Cas9 to the target DNA the target DNA via Watson-Crick base-pairing between the guide sequence portion of the crRNA and the protospacer on the target DNA next to the protospacer adjacent motif (PAM). A skilled artisan will appreciate that each of the engineered RNA molecule of the present invention is further designed such as to associate with a target genomic DNA sequence of interest next to a protospacer adjacent motif (PAM), e.g., a PAM matching the sequence relevant for the type of CRISPR nuclease utilized, such as for a non-limiting example, NGG or NAG, wherein “N” is any nucleobase, for Streptococcus pyogenes Cas9 WT (SpCAS9); NNGRRT for Staphylococcus aureus (SaCas9); NNNVRYM for Jejuni Cas9 WT; NGAN or NGNG for SpCas9-VQR variant; NGCG for SpCas9-VRER variant; NGAG for SpCas9-EQR variant; NRRH for SpCas9-NRRH variant, wherein N is any nucleobase, R is A or G and H is A, C, or T; NRTH for SpCas9-NRTH variant, wherein N is any nucleobase, R is A or G and H is A, C, or T; NRCH for SpCas9-NRCH variant, wherein N is any nucleobase, R is A or G and H is A, C, or T; NG for SpG variant of SpCas9 wherein N is any nucleobase; NG or NA for SpCas9-NG variant of SpCas9 wherein N is any nucleobase; NR or NRN or NYN for SpRY variant of SpCas9, wherein N is any nucleobase, R is A or G and Y is C or T; NNG for Streptococcus canis Cas9 variant (ScCas9), wherein N is any nucleobase; NNNRRT for SaKKH-Cas9 variant of Staphylococcus aureus (SaCas9), wherein N is any nucleobase, and R is A or G; NNNNGATT for Neisseria meningitidis (NmCas9), wherein N is any nucleobase; TTN for Alicyclobacillus acidiphilus Cas12b (AacCas12b), wherein N is any nucleobase; or TTTV for Cpf1, wherein V is A, C or G. RNA molecules of the present invention are each designed to form complexes in conjunction with one or more different CRISPR nucleases and designed to target polynucleotide sequences of interest utilizing one or more different PAM sequences respective to the CRISPR nuclease utilized.

In some embodiments, an RNA-guided DNA nuclease e.g., a CRISPR nuclease, may be used to cause a DNA break, either double or single-stranded in nature, at a desired location in the genome of a cell. The most commonly used RNA-guided DNA nucleases are derived from CRISPR systems, however, other RNA-guided DNA nucleases are also contemplated for use in the genome editing compositions and methods described herein. For instance, see U.S. Patent Publication No. 2015/0211023, incorporated herein by reference.

CRISPR systems that may be used in the practice of the invention vary greatly. CRISPR systems can be a type I, a type II, or a type III system. Non-limiting examples of suitable CRISPR proteins include Cas3, Cas4, Cas5, Cas5e (or CasD), Cas6, Cas6e, Cas6f, Cas7, Cas8a1, Cas8a2, Cas8b, Cas8c, Cas9, Cas10, Casl Od, CasF, CasG, CasH, Csy1, Csy2, Csy3, Cse1 (or CasA), Cse2 (or CasB), Cse3 (or CasE), Cse4 (or CasC), Cscl, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3,Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Cszl, Csx5, Csf1, Csf2, Csf3, Csf4, and Cul966.

In some embodiments, the RNA-guided DNA nuclease is a CRISPR nuclease derived from a type II CRISPR system (e.g., Cas9). The CRISPR nuclease may be derived from Streptococcus pyogenes, Streptococcus thermophilus, Streptococcus sp., Staphylococcus aureus, Neisseria meningitidis, Treponema denticola, Nocardiopsis dassonvillei, Streptomyces pristinaespiralis, Streptomyces viridochromogenes, Streptomyces viridochromogenes, Streptosporangium roseum, Streptosporangium roseum, Alicyclobacillus acidocaldarius, Bacillus pseudomycoides, Bacillus selenitireducens, Exiguobacterium sibiricum, Lactobacillus delbrueckii, Lactobacillus salivarius, Microscilla marina, Burkholderiales bacterium, Polaromonas naphthalenivorans, Polaromonas sp., Crocosphaera watsonii, Cyanothece sp., Microcystis aeruginosa, Synechococcus sp., Acetohalobium arabaticum, Ammonifex degensii, Caldicelulosiruptor becscii, Candidatus Desulforudis, Clostridium botulinum, Clostridium Finegoldia magna, Natranaerobius thermophilus, Pelotomaculumthermopropionicum, Acidithiobacillus caldus, Acidithiobacillus ferrooxidans, Allochromatium vinosum, Marinobacter sp., Nitrosococcus halophilus, Nitrosococcus watsoni, Pseudoalteromonas haloplanktis, Ktedonobacter racemifer, Methanohalobium evestigatum, Anabaena variabilis, Nodularia spumigena, Nostoc sp., Arthrospira maxima, Arthrospira platensis, Arthrospira sp., Lyngbya sp., Microcoleus chthonoplastes, Oscillatoria sp., Petrotoga mobilis, Thermosipho africanus, Acaryochloris marina, or any species which encodes a CRISPR nuclease with a known PAM sequence. CRISPR nucleases encoded by uncultured bacteria may also be used in the context of the invention. (See Burstein et al. Nature, 2017). Variants of CRIPSR proteins having known PAM sequences e.g., SpCas9 D1135E variant, SpCas9 VQR variant, SpCas9 EQR variant, or SpCas9 VRER variant may also be used in the context of the invention.

Thus, an RNA guided DNA nuclease of a CRISPR system, such as a Cas9 protein or modified Cas9 or homolog or ortholog of Cas9, or other RNA guided DNA nucleases belonging to other types of CRISPR systems, such as Cpf1 and its homologs and orthologs, may be used in the compositions of the present invention. Additional CRISPR nucleases may also be used, for example, the nucleases described in PCT International Application Publication Nos. WO2020/223514 and WO2020/223553, which are hereby incorporated by reference.

In certain embodiments, the CRIPSR nuclease may be a “functional derivative” of a naturally occurring Cas protein. A “functional derivative” of a native sequence polypeptide is a compound having a qualitative biological property in common with a native sequence polypeptide. “Functional derivatives” include, but are not limited to, fragments of a native sequence and derivatives of a native sequence polypeptide and its fragments, provided that they have a biological activity in common with a corresponding native sequence polypeptide. A biological activity contemplated herein is the ability of the functional derivative to hydrolyze a DNA substrate into fragments. The term “derivative” encompasses both amino acid sequence variants of polypeptide, covalent modifications, and fusions thereof. Suitable derivatives of a Cas polypeptide or a fragment thereof include but are not limited to mutants, fusions, covalent modifications of Cas protein or a fragment thereof. Cas protein, which includes Cas protein or a fragment thereof, as well as derivatives of Cas protein or a fragment thereof, may be obtainable from a cell or synthesized chemically or by a combination of these two procedures. The cell may be a cell that naturally produces Cas protein, or a cell that naturally produces Cas protein and is genetically engineered to produce the endogenous Cas protein at a higher expression level or to produce a Cas protein from an exogenously introduced nucleic acid, which nucleic acid encodes a Cas that is same or different from the endogenous Cas. In some cases, the cell does not naturally produce Cas protein and is genetically engineered to produce a Cas protein.

In some embodiments, the CRISPR nuclease is Cpf1. Cpf1 is a single RNA-guided endonuclease which utilizes a T-rich protospacer-adjacent motif. Cpf1 cleaves DNA via a staggered DNA double-stranded break. Two Cpf1 enzymes from Acidaminococcus and Lachnospiraceae have been shown to carry out efficient genome-editing activity in human cells. (See Zetsche et al., 2015).

Thus, an RNA guided DNA nuclease of a Type II CRISPR System, such as a Cas9 protein or modified Cas9 or homologs, orthologues, or variants of Cas9, or other RNA guided DNA nucleases belonging to other types of CRISPR systems, such as Cpf1 and its homologs, orthologues, or variants, may be used in the present invention.

In some embodiments, the guide molecule comprises one or more chemical modifications which imparts a new or improved property (e.g., improved stability from degradation, improved hybridization energetics, or improved binding properties with an RNA guided DNA nuclease). Suitable chemical modifications include, but are not limited to: modified bases, modified sugar moieties, or modified inter-nucleoside linkages. Non-limiting examples of suitable chemical modifications include: 4-acetylcytidine, 5-(carboxyhydroxymethyl)uridine, 2′-O-methylcytidine, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluridine, dihydrouridine, 2′-O-methylpseundouridine, “beta, D-galactosylquenosine”, 2′-O-methylguanosine, inosine, N6-isopentenyladenosine. 1-methyladenosine, 1-methylpseudouridine, 1-methylguanosine, 1-methylinosine, “2,2-dimethylguanosine”, 2-methyladenosine, 2-methylguanosine, 3-methylcytidine, 5-methylcytidine, N6-methyladenosine, 7-methylguanosine, 5-methylaminomethyluridine, 5-methoxyaminomethyl-2-thiouridine, “beta, D-mannosylqueuosine”, 5-methoxycarbonylmethyl-2-thiouridine, 5-methoxycarbonylmethyluridine, 5-methoxyuridine, 2-methylthio-N6-isopentenyladenosine, N-((9-beta-D-ribofuranosyl-2-methylthiopurine-6-yl)carbamoyl)threonine, N4(9-beta-D-ribofuranosylpurine-6-yl)N-methylcarbamoyl)threonine, uridine-5-oxyacetic acid-methylester, uridine-5-oxyacetic acid, wybutoxosine, queuosine, 2-thiocytidine, 5-methyl-2-thiouridine, 2-thiouridine, 4-thiouridine, N-((9-beta-D-ribofuranosylpurine-6-yl)-carbamoyl)threonine, 2′-O-methyl-2′-O-methyluridine, wybutosine, “3-(3-amino-3-carboxy-propyl)uridine, (acp3)u”, 2′-0-methyl (M), 3′-phosphorothioate (MS), 3′-thioPACE (MSP), pseudouridine, or 1-methyl pseudo-uridine. Each possibility represents a separate embodiment of the present invention.

Guide Sequences which Specifically Target a Mutant Allele

A given gene may contain thousands of SNPs. Utilizing a twenty-five base pair target window for targeting each SNP in a gene would require hundreds of thousands of guide sequences. Any given guide sequence when utilized to target a SNP may result in degradation of the guide sequence, limited activity, no activity, or off-target effects. Accordingly, suitable guide sequences are necessary for targeting a given gene. By the present invention, a novel set of guide sequences have been identified for knocking out expression of a mutated LRRK2 protein, inactivating a mutant LRRK2 gene allele, and treating monogenic Parkinson's disease.

The present disclosure provides guide sequences capable of specifically targeting a mutated allele for inactivation while leaving the functional allele unmodified. The guide sequences of the present invention are designed to, and are most likely to, specifically differentiate between a mutated allele and a functional allele. Of all possible guide sequences which target a mutated allele desired to be inactivated, the specific guide sequences disclosed herein are specifically effective to function with the disclosed embodiments.

Briefly, the guide sequences may have properties as follows: (1) target SNP/insertion/deletion/indel with a high prevalence in the general population, in a specific ethnic population or in a patient population is above 1% and the SNP/insertion/deletion/indel heterozygosity rate in the same population is above 1%; (2) target a location of a SNP/insertion/deletion/indel proximal to a portion of the gene e.g., within 5k bases of any portion of the gene, for example, a promoter, a UTR, an exon or an intron; and (3) target a mutant allele using an RNA molecule which targets a founder or common pathogenic mutations for the disease/gene. In some embodiments, the prevalence of the SNP/insertion/deletion/indel in the general population, in a specific ethnic population or in a patient population is above 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15% and the SNP/insertion/deletion/indel heterozygosity rate in the same population is above 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15%. Each possibility represents a separate embodiment and may be combined at will.

For each gene, according to SNP/insertion/deletion/indel any one of the following strategies may be used to deactivate the mutated allele: (1) Knockout strategy using one RNA molecule—one RNA molecule is utilized to direct a CRISPR nuclease to a mutated allele and create a double-strand break (DSB) leading to formation of a frameshift mutation in an exon or in a splice site region of the mutated allele; and (2) Excision of at least one coding exon or a complete knockout of a mutant LRRK2 allele using two RNA molecules, for example, a first RNA molecule targets a SNP position of an Intron 2 of the mutant LRRK2 allele and a second, non-discriminatory RNA molecule targets a sequence in Intron 1, preferably within 500 basepairs upstream to Exon 2, of the LRRK2 gene.

Based on the locations of identified SNPs/insertions/deletions/indels for each mutant allele, any one of, or a combination of, the above-mentioned methods to deactivate the mutant allele may be utilized.

In some embodiments of the present invention, an RNA molecule is used to target a pathogenic mutation within a mutant LRRK2 allele. In some embodiments of the present invention, an RNA molecule is used to target a SNP position. In some embodiments, the location of the SNP position is within a region of three kilobases upstream of the transcriptional start site to three kilobases downstream of the LRRK2 3′UTR.

Guide sequences of the present invention may: (1) target a heterozygous SNP for the targeted gene; (2) target a heterozygous SNP upstream or downstream of the gene; (3) target a SNP with a prevalence of the SNP/insertion/deletion/indel in the general population, in a specific ethnic population, or in a patient population above 1%; (4) have a guanine-cytosine content of greater than 30% and less than 85%; (5) have no repeat of seven or more thymine/uracil, guanine, cytosine, or adenine; and (6) have no additional target in the genome with zero mismatch with the same PAM sequence. Guide sequences of the present invention may satisfy any one of the above criteria and are most likely to differentiate between a mutated allele from its corresponding functional allele.

In some embodiments of the present invention, at least one nucleotide which differs between the mutated allele and the functional allele is upstream, downstream or within the sequence of the disease-causing mutation of the gene of interest. The at least one nucleotide which differs between the mutated allele and the functional allele may be within an exon or within an intron of the gene of interest. In some embodiments, the at least one nucleotide which differs between the mutated allele and the functional allele is within an exon of the gene of interest. In some embodiments, the at least one nucleotide which differs between the mutated allele and the functional allele is within an intron or the exon of the gene of interest, in close proximity to the splice site between the intron and the exon e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides upstream or downstream to the splice site. Each possibility represents a separate embodiment.

In some embodiments, the at least one nucleotide is a single nucleotide polymorphism (SNP). In some embodiments, each of the nucleotide variants of the SNP may be expressed in the mutated allele. In some embodiments, the SNP may be a founder or common pathogenic mutation.

Guide sequences may target a SNP which has both (1) a high prevalence in the general population e.g., above 1% in the population; and (2) a high heterozygosity rate in the population, e.g., above 1%. Guide sequences may target a SNP that is globally distributed. A SNP may be a founder or common pathogenic mutation. In some embodiments, the prevalence in the general population is above 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15%. Each possibility represents a separate embodiment. In some embodiments, the heterozygosity rate in the population is above 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15%. Each possibility represents a separate embodiment.

In some embodiments, the at least one nucleotide which differs between the mutated allele and the functional allele is linked to/co-exists with the disease-causing mutation in high prevalence in a population. In such embodiments, “high prevalence” refers to at least 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. Each possibility represents a separate embodiment of the present invention. In one embodiment, the at least one nucleotide which differs between the mutated allele and the functional allele, is a disease-associated mutation. In some embodiments, the SNP is highly prevalent in the population. In such embodiments, “highly prevalent” refers to at least 10%, 11%, 12%, 13%, 14%, 15%, 20%, 30%, 40%, 50%, 60%, or 70% of a population. Each possibility represents a separate embodiment of the present invention.

Delivery to Cells

The RNA molecule compositions described herein may be delivered to a target cell by any suitable means. RNA molecule compositions of the present invention may be targeted to any cell which contains and/or expresses a mutated allele, including any mammalian or plant cell. For example, in one embodiment the RNA molecule specifically targets a mutated LRRK2 allele and the target cell is an HSC. The delivery to the cell may be performed in-vitro, ex-vivo, or in-vivo. Further, the nucleic acid compositions described herein may be delivered as one or more of DNA molecules, RNA molecules, ribonucleoproteins (RNPs), nucleic acid vectors, or any combination thereof.

In some embodiments, any one of the compositions described herein is in the form of an RNP composition and is delivered to a cell ex-vivo. In some embodiments, the cell is a stem cell, a neuron, or a dopaminergic cell. The RNP composition may be delivered to the cell by any known ex-vivo delivery method, including but not limited to, electroporation, viral transduction, nanoparticle delivery, liposomes, etc. Additional detailed delivery methods are described throughout this section.

In some embodiments, the RNA molecule comprises a chemical modification. Non-limiting examples of suitable chemical modifications include 2′-0-methyl (M), 2′-0-methyl, 3′phosphorothioate (MS) or 2′-0-methyl, 3′thioPACE (MSP), pseudouridine, and 1-methyl pseudo-uridine. Each possibility represents a separate embodiment of the present invention.

Any suitable viral vector system may be used to deliver nucleic acid compositions e.g., the RNA molecule compositions of the subject invention. Conventional viral and non-viral based gene transfer methods can be used to introduce nucleic acids and target tissues. In certain embodiments, nucleic acids are administered for in vivo or ex vivo gene therapy uses. Non-viral vector delivery systems include naked nucleic acid, and nucleic acid complexed with a delivery vehicle such as a liposome or poloxamer. For a review of gene therapy procedures, see Anderson (1992); Nabel & Felgner (1993); Mitani & Caskey (1993); Dillon (1993); Miller (1992); Van Brunt (1988); Vigne (1995); Kremer & Perricaudet (1995); Haddada et al. (1995); and Yu et al. (1994).

Methods of non-viral delivery of nucleic acids and/or proteins include electroporation, lipofection, microinjection, biolistics, particle gun acceleration, virosomes, liposomes, immunoliposomes, lipid nanoparticles (LNPs), polycation or lipid:nucleic acid conjugates, artificial virions, and agent-enhanced uptake of nucleic acids or can be delivered to plant cells by bacteria or viruses (e.g., Agrobacterium, Rhizobium sp. NGR234, Sinorhizoboiummeliloti, Mesorhizobium loti, tobacco mosaic virus, potato virus X, cauliflower mosaic virus and cassava vein mosaic virus). (See, e.g., Chung et al., 2006). Sonoporation using, e.g., the Sonitron 2000 system (Rich-Mar), can also be used for delivery of nucleic acids. Cationic-lipid mediated delivery of proteins and/or nucleic acids is also contemplated as an in vivo, ex vivo, or in vitro delivery method. (See Zuris et al. (2015); see also Coelho et al. (2013); Judge et al. (2006); and Basha et al. (2011)).

Non-viral vectors, such as transposon-based systems e.g. recombinant Sleeping Beauty transposon systems or recombinant PiggyBac transposon systems, may also be delivered to a target cell and utilized for transposition of a polynucleotide sequence of a molecule of the composition or a polynucleotide sequence encoding a molecule of the composition in the target cell.

Additional exemplary nucleic acid delivery systems include those provided by Amaxa.RTM. Biosystems (Cologne, Germany), Maxcyte, Inc. (Rockville, Md.), BTX Molecular Delivery Systems (Holliston, Mass.) and Copernicus Therapeutics Inc., (see, e.g., U.S. Pat. No. 6,008,336). Lipofection is described in e.g., U.S. Pat. Nos. 5,049,386, 4,946,787; and 4,897,355, and lipofection reagents are sold commercially (e.g., Transfectam.™., Lipofectin.™. and Lipofectamine.™. RNAiMAX). Cationic and neutral lipids that are suitable for efficient receptor-recognition lipofection of polynucleotides include those disclosed in PCT International Publication Nos. WO/1991/017424 and WO/1991/016024. Delivery can be to cells (ex vivo administration) or target tissues (in vivo administration).

The preparation of lipid:nucleic acid complexes, including targeted liposomes such as immunolipid complexes, is well known to one of skill in the art (see, e.g., Crystal, Science (1995); Blaese et al., (1995); Behr et al., (1994); Remy et al. (1994); Gao and Huang (1995); Ahmad and Allen (1992); U.S. Pat. Nos. 4,186,183; 4,217,344; 4,235,871; 4,261,975; 4,485,054; 4,501,728; 4,774,085; 4,837,028; and 4,946,787).

Additional methods of delivery include the use of packaging the nucleic acids to be delivered into EnGeneIC delivery vehicles (EDVs). These EDVs are specifically delivered to target tissues using bispecific antibodies where one arm of the antibody has specificity for the target tissue and the other has specificity for the EDV. The antibody brings the EDVs to the target cell surface and then the EDV is brought into the cell by endocytosis. Once in the cell, the contents are released (See MacDiarmid et al., 2009).

The use of RNA or DNA viral based systems for viral mediated delivery of nucleic acids take advantage of highly evolved processes for targeting a virus to specific cells in the body and trafficking the viral payload to the nucleus. Viral vectors can be administered directly to patients (in vivo) or they can be used to treat cells in vitro and the modified cells are administered to patients (ex vivo). Conventional viral based systems for the delivery of nucleic acids include, but are not limited to, retroviral, lentivirus, adenoviral, adeno-associated, vaccinia and herpes simplex virus vectors for gene transfer.

The tropism of a retrovirus can be altered by incorporating foreign envelope proteins, expanding the potential target population of target cells. Lentiviral vectors are retroviral vectors that are able to transduce or infect non-dividing cells and typically produce high viral titers. Selection of a retroviral gene transfer system depends on the target tissue. Retroviral vectors are comprised of cis-acting long terminal repeats with packaging capacity for up to 6-10 kb of foreign sequence. The minimum cis-acting LTRs are sufficient for replication and packaging of the vectors, which are then used to integrate the therapeutic gene into the target cell to provide permanent transgene expression. Widely used retroviral vectors include those based upon murine leukemia virus (MuLV), gibbon ape leukemia virus (GaLV), Simian Immunodeficiency virus (SIV), human immunodeficiency virus (HIV), and combinations thereof (See, e.g., Buchschacher et al. (1992); Johann et al. (1992); Sommerfelt et al. (1990); Wilson et al. (1989); Miller et al. (1991); PCT International Publication No. WO/1994/026877A1).

At least six viral vector approaches are currently available for gene transfer in clinical trials, which utilize approaches that involve complementation of defective vectors by genes inserted into helper cell lines to generate the transducing agent.

pLASN and MFG-S are examples of retroviral vectors that have been used in clinical trials (See Dunbar et al., 1995; Kohn et al., 1995; Malech et al., 1997). PA317/pLASN was the first therapeutic vector used in a gene therapy trial (Blaese et al., 1995). Transduction efficiencies of 50% or greater have been observed for MFG-S packaged vectors. (Ellem et al., (1997); Dranoff et al., 1997).

Packaging cells are used to form virus particles that are capable of infecting a host cell. Such cells include 293 cells, which package adenovirus, AAV, and Psi-2 cells or PA317 cells, which package retrovirus. Viral vectors used in gene therapy are usually generated by a producer cell line that packages a nucleic acid vector into a viral particle. The vectors typically contain the minimal viral sequences required for packaging and subsequent integration into a host (if applicable), other viral sequences being replaced by an expression cassette encoding the protein to be expressed. The missing viral functions are supplied in trans by the packaging cell line. For example, AAV vectors used in gene therapy typically only possess inverted terminal repeat (ITR) sequences from the AAV genome which are required for packaging and integration into the host genome. Viral DNA is packaged in a cell line, which contains a helper plasmid encoding the other AAV genes, namely rep and cap, but lacking ITR sequences. The cell line is also infected with adenovirus as a helper. The helper virus promotes replication of the AAV vector and expression of AAV genes from the helper plasmid. The helper plasmid is not packaged in significant amounts due to a lack of ITR sequences. Contamination with adenovirus can be reduced by, e.g., heat treatment to which adenovirus is more sensitive than AAV. Additionally, AAV can be produced at clinical scale using baculovirus systems (see U.S. Pat. No. 7,479,554).

In many gene therapy applications, it is desirable that the gene therapy vector be delivered with a high degree of specificity to a particular tissue type. Accordingly, a viral vector can be modified to have specificity for a given cell type by expressing a ligand as a fusion protein with a viral coat protein on the outer surface of the virus. The ligand is chosen to have affinity for a receptor known to be present on the cell type of interest. For example, Han et al. (1995) reported that Moloney murine leukemia virus can be modified to express human heregulin fused to gp70, and the recombinant virus infects certain human breast cancer cells expressing human epidermal growth factor receptor. This principle can be extended to other virus-target cell pairs, in which the target cell expresses a receptor and the virus expresses a fusion protein comprising a ligand for the cell-surface receptor. For example, filamentous phage can be engineered to display antibody fragments (e.g., FAB or Fv) having specific binding affinity for virtually any chosen cellular receptor. Although the above description applies primarily to viral vectors, the same principles can be applied to nonviral vectors. Such vectors can be engineered to contain specific uptake sequences which favor uptake by specific target cells.

Gene therapy vectors can be delivered in vivo by administration to an individual patient, typically by systemic administration (e.g., intravitreal, intravenous, intraperitoneal, intramuscular, subdermal, or intracranial infusion) or topical application, as described below. Alternatively, vectors can be delivered to cells ex vivo, such as cells explanted from an individual patient (e.g., lymphocytes, bone marrow aspirates, tissue biopsy) or universal donor hematopoietic stem cells, followed by reimplantation of the cells into a patient, optionally after selection for cells which have incorporated the vector. A non-limiting exemplary ex vivo approach may involve removal of tissue (e.g., peripheral blood, bone marrow, and spleen) from a patient for culture, nucleic acid transfer to the cultured cells (e.g., hematopoietic stem cells), followed by grafting the cells to a target tissue (e.g., bone marrow, and spleen) of the patient. In some embodiments, the stem cell or hematopoietic stem cell may be further treated with a viability enhancer.

Ex vivo cell transfection for diagnostics, research, or for gene therapy (e.g., via re-infusion of the transfected cells into the host organism) is well known to those of skill in the art. In a preferred embodiment, cells are isolated from the subject organism, transfected with a nucleic acid composition, and re-infused back into the subject organism (e.g., patient). Various cell types suitable for ex vivo transfection are well known to those of skill in the art (See, e.g., Freshney, “Culture of Animal Cells, A Manual of Basic Technique and Specialized Applications (6th edition, 2010) and the references cited therein for a discussion of how to isolate and culture cells from patients).

Suitable cells include, but are not limited to, eukaryotic cells and/or cell lines. Non-limiting examples of such cells or cell lines generated from such cells include COS, CHO (e.g., CHO—S, CHO-K1, CHO-DG44, CHO-DUXB11, CHO-DUKX, CHOK1SV), VERO, MDCK, WI38, V79, B14AF28-G3, BHK, HaK, NSO, SP2/0-Ag14, HeLa, HEK293 (e.g., HEK293-F, HEK293-H, HEK293-T), perC6 cells, any plant cell (differentiated or undifferentiated), as well as insect cells such as Spodopterafugiperda (Sf), or fungal cells such as Saccharomyces, Pichia and Schizosaccharomyces. In certain embodiments, the cell line is a CHO-K1, MDCK or HEK293 cell line. Additionally, primary cells may be isolated and used ex vivo for reintroduction into the subject to be treated following treatment with a guided nuclease system (e.g. CRISPR/Cas). Suitable primary cells include peripheral blood mononuclear cells (PBMC), and other blood cell subsets such as, but not limited to, CD4+ T cells or CD8+ T cells. Suitable cells also include stem cells such as, by way of example, embryonic stem cells, induced pluripotent stem cells, hematopoietic stem cells (CD34+), neuronal stem cells and mesenchymal stem cells.

In one embodiment, stem cells are used in ex vivo procedures for cell transfection and gene therapy. The advantage to using stem cells is that they can be differentiated into other cell types in vitro, or can be introduced into a mammal (such as the donor of the cells) where they will engraft in the bone marrow. Methods for differentiating CD34+ cells in vitro into clinically important immune cell types using cytokines such a GM-CSF, IFN-gamma, and TNF-alpha are known (as a non-limiting example see, Inaba et al., 1992).

Stem cells are isolated for transduction and differentiation using known methods. For example, stem cells are isolated from bone marrow cells by panning the bone marrow cells with antibodies which bind unwanted cells, such as CD4+ and CD8+ (T cells), CD45+(panB cells), GR-1 (granulocytes), and Tad (differentiated antigen presenting cells) (as a non-limiting example, see Inaba et al., 1992). Stem cells that have been modified may also be used in some embodiments.

Vectors (e.g., retroviruses, liposomes, etc.) containing therapeutic nucleic acid compositions can also be administered directly to an organism for transduction of cells in vivo. Administration is by any of the routes normally used for introducing a molecule into ultimate contact with blood or tissue cells including, but not limited to, injection, infusion, topical application (e.g., eye drops and cream) and electroporation. Suitable methods of administering such nucleic acids are available and well known to those of skill in the art, and, although more than one route can be used to administer a particular composition, a particular route can often provide a more immediate and more effective reaction than another route. According to some embodiments, the composition is delivered via IV injection.

Vectors suitable for introduction of transgenes into immune cells (e.g., T-cells) include non-integrating lentivirus vectors. See, e.g., U.S. Patent Publication No. 2009/0117617.

Pharmaceutically acceptable carriers are determined in part by the particular composition being administered, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of pharmaceutical compositions available, as described below (See, e.g., Remington's Pharmaceutical Sciences, 17th ed., 1989).

In accordance with some embodiments, there is provided an RNA molecule which binds to/associates with and/or directs the RNA guided DNA nuclease to a sequence comprising at least one nucleotide which differs between a mutated allele and a functional allele (e.g., SNP) of a gene of interest (i.e., a sequence of the mutated allele which is not present in the functional allele). The sequence may be within the disease associated mutation. The sequence may be upstream or downstream to the disease associated mutation. Any sequence difference between the mutated allele and the functional allele may be targeted by an RNA molecule of the present invention to inactivate the mutant allele, or otherwise disable its dominant disease-causing effects, while preserving the activity of the functional allele.

The disclosed compositions and methods may also be used in the manufacture of a medicament for treating dominant genetic disorders in a patient.

Mechanisms of Action for LRRK2 Knockout Methods

Without being bound by any theory or mechanism, the instant invention may be utilized to apply a CRISPR nuclease to process a mutated pathogenic LRRK2 allele and not a functional LRRK2 allele, such as to prevent expression of the mutated pathogenic allele or to produce a truncated non-pathogenic peptide from the mutated pathogenic allele, in order to prevent or treat monogenic Parkinson's disease. A specific guide sequence may be selected from Table 1 based on the targeted SNP position and the type of CRISPR nuclease used (e.g. according to a required PAM sequence). Notably, according to the Genome Aggregation Database (gnomAd) there are healthy individuals with loss of function mutations in one of their LRRK2 alleles. Therefore, there is no concern for haploinsufficiency upon knocking out a mutated LRRK2 allele.

The LRRK2 gene is located on chromosome 12 and encodes the Leucine-rich repeat serine/threonine-protein kinase 2 protein. One strategy to knockout a LRRK2 mutant allele is to target a pathogenic mutation position using one RNA molecule in order to mediate truncation or nonsense mediated decay (NMD) of the LRRK2 mutant allele. As a non-limiting example, a frameshift in a mutated LRRK2 allele may be introduced by utilizing one RNA molecule to target a pathogenic mutation or SNP position in a coding exon of the mutated LRRK2 allele (e.g. rs10878245 in Exon 5, rs1427263 in Exon 34 and rs10878405 in Exon 43) in order to mediate a double-strand break, which leads to generation of a frameshift mutation and expression of a truncated protein or nonsense mediated decay (NMD) of its transcripts.

Alternatively, a LRRK2 mutant allele may be knocked-out by an excision strategy that utilizes two RNA molecules. In such strategies, SNP positions within three kilobases upstream to the transcription start site and three kilobases downstream of the 3′UTR of the LRRK2 gene may be targeted.

In one excision strategy approach, Exon 2 of a mutant LRRK2 allele is excised by utilizing a first RNA molecule to target a SNP position in Intron 2 of the mutant LRRK2 allele and utilizing a second non-discriminatory RNA molecule to target a region in Intron 1, of the LRRK2 gene. In some embodiments of this approach, the target region is within 500 bps of Exon 2.

In another excision strategy approach, Exon 3 of a mutant LRRK2 allele is excised by utilizing a first RNA molecule to target a SNP position in Intron 4 and utilizing a second non-discriminatory RNA molecule to target a region in Intron 3. In some embodiments of this approach, the target region is within 500 bps of Exon 4.

In another excision strategy approach, the 3′UTR of a mutant LRRK2 allele is excised by utilizing a first RNA molecule to target a SNP position in Intron 49 and utilizing a second non-discriminatory RNA molecule to target a region within 500 base pairs downstream to Exon 51. In some embodiments of this approach, the target region is within 500 bps of Exon 4.

Alternatively, the 3′UTR of a mutant LRRK2 allele is excised by utilizing a first RNA molecule to target a SNP position downstream of Exon 51 and utilizing a second non-discriminatory RNA molecule to target a region in Intron 50 or Intron 49, preferably within 500 base pairs upstream to Exon 50.

In another excision strategy approach, a promoter region of a mutant LRRK2 allele is excised by utilizing a first RNA molecule to target a SNP with 3000 base pairs of the transcriptional start site and utilizing a second non-discriminatory RNA molecule to target a region in Intron 1 or Intron 2, preferably within 500 base pairs downstream to Exon 2.

Examples of RNA Guide Sequences which Specifically Target Mutated Alleles of LRRK2 Gene

Although a large number of guide sequences can be designed to target a mutated allele, the nucleotide sequences described in Table 1 identified by SEQ ID NOs: 1-103,606 below were specifically selected to effectively implement the methods set forth herein and to effectively discriminate between alleles.

Table 1 shows guide sequences designed for use as described in the embodiments above to associate with different SNPs or pathogenic mutations within a sequence of a mutated LRRK2 allele. Each engineered guide molecule is further designed such as to associate with a target genomic DNA sequence of interest that lies next to a protospacer adjacent motif (PAM), e.g., a PAM matching the sequence NGG or NAG, where “N” is any nucleobase. The guide sequences were designed to work in conjunction with one or more different CRISPR nucleases, including, but not limited to, e.g. SpCas9WT (PAM SEQ: NGG), SpCas9.VQR.1 (PAM SEQ: NGAN), SpCas9.VQR.2 (PAM SEQ: NGNG), SpCas9.EQR (PAM SEQ: NGAG), SpCas9.VRER (PAM SEQ: NGCG), SaCas9WT (PAM SEQ: NNGRRT), SpRY (PAM SEQ: NRN or NYN), NmCas9WT (PAM SEQ: NNNNGATT), Cpf1 (PAM SEQ: TTTV), or JeCas9WT (PAM SEQ: NNNVRYM). RNA molecules of the present invention are each designed to form complexes in conjunction with one or more different CRISPR nucleases and designed to target polynucleotide sequences of interest utilizing one or more different PAM sequences respective to the CRISPR nuclease utilized.

TABLE 1 Guide sequences designed to associate with specific LRRK2 gene targets SEQ ID NOS: of 20 base SEQ ID NOs: of SEQ ID NOs: of Target guides 21 base guides 22 base guides 12:40225041_C_T 1-46 47-96 97-148 12:40225058_C_A 149-196 197-246 247-298 12:40225088_G_A 299-345 346-395 396-447 12:40225159_G_A 448-495 496-545 546-597 12:40225233_G_C 598-645 646-695 696-747 12:40225558_C_T 748-791 792-833 834-877 12:40225618_T_C 878-921 922-971 972-1023 12:40225628_G_A 1024-1070 1071-1120 1121-1172 12:40225631_T_C 1024, 1035, 1072, 1083, 1121, 1123, 1049, 1055, 1104, 1116, 1168, 1171, 1173-1215 1216-1261 1262-1309 12:40232303_A_C 1310-1356 1357-1406 1407-1458 12:40232345_G_C 1459-1505 1506-1555 1556-1607 12:40232380_A_C 1608-1652 1653-1698 1699-1746 12:40232385_T_C 1747-1788 1789-1828 1829-1870 12:40235634_T_C 1871-1912 1913-1945 1946-1988 12:40235646_C_G 1989-2018 2019-2041 2042-2068 12:40235660_A_G 2069-2113 2114-2160 2161-2209 12:40238078_A_G 2210-2257 2258-2307 2308-2359 12:40240517_C_T 2360-2367 2368-2373 2374-2381 12:40240531_T_C 2382-2400 2401-2415 2416-2432 12:40240594_G_C 2433-2479 2480-2529 2530-2581 12:40240602_T_C 2582-2629 2630-2679 2680-2731 12:40243551_CAAT_C 2732-2767 2768-2808 2809-2852 12:40243556_A_T 2738, 2752, 2776, 2784, 2818, 2833, 2760, 2763, 2791, 2793, 2835, 2839, 2853-2885 2803-2804, 2846-2847, 2886-2918 2851, 2919-2953 12:40249822_T_G 2954-2973 2974-2982 2983-2997 12:40249843_C_G 2998-3019 3020-3039 3040-3065 12:40249854_C_T 3066-3098 3099-3126 3127-3158 12:40249881_T_C 3159-3205 3206-3255 3256-3307 12:40249908_A_G 3308-3355 3356-3405 3406-3457 12:40251273_G_A 3458-3475 3476-3480 3481-3490 12:40251303_G_A 3491-3526 3527-3563 3564-3603 12:40251361_A_G 3604-3651 3652-3701 3702-3753 12:40251366_C_A 3754-3800 3801-3850 3851-3902 12:40251369_G_A 3761, 3775, 3803, 3808, 3853, 3868, 3799, 3903- 3818, 3850, 3890, 3894, 3945 3946-3991 3992-4039 12:40251495_C_T 4040-4065 4066-4083 4084-4108 12:40252906_A_G 4109-4135 4136-4163 4164-4195 12:40252929_G_A 4196-4243 4244-4293 4294-4345 12:40252939_C_T 4346-4392 4393-4442 4443-4494 12:40252952_T_G 4495-4541 4542-4591 4592-4643 12:40252958_A_C 4644-4691 4692-4741 4742-4793 12:40252968_G_A 4794-4834 4835-4874 4875-4920 12:40252974_C_T 4921-4966 4967-5012 5013-5063 12:40252978_C_T 4944, 4964, 4998, 5004, 5020, 5060, 5064-5108 5109-5156 5157-5206 12:40252984_C_T 5207-5253 5254-5303 5304-5355 12:40252988_T_C 5236, 5244, 5262, 5300, 5306, 5338, 5356-5400 5401-5448 5449-5498 12:40253016_G_A 5499-5537 5538-5567 5568-5605 12:40257311_A_G 5606-5648 5649-5698 5699-5750 12:40257341_G_T 5751-5796 5797-5838 5839-5884 12:40259507_A_G 5885-5932 5933-5982 5983-6034 12:40259525_A_T 6035-6074 6075-6112 6113-6154 12:40259534_G_T 6155-6186 6187-6211 6212-6243 12:40259578_G_A 6244-6277 6278-6311 6312-6347 12:40263806_A_G 6348-6394 6395-6442 6443-6493 12:40263875_A_G 6494-6538 6539-6579 6580-6626 12:40263900_G_A 6627-6662 6663-6690 6691-6720 12:40263907_T_C 6721-6756 6757-6788 6789-6824 12:40274646_CC_C 6825-6858 6859-6879 6880-6908 12:40274881_T_C 6909-6953 6954-6993 6994-7040 12:40274899_A_G 7041-7084 7085-7124 7125-7169 12:40274917_G_C 7170-7216 7217-7262 7263-7311 12:40277933_T_C 7312-7359 7360-7408 7409-7460 12:40277968_A_C 7461-7500 7501-7523 7524-7559 12:40277971_A_G 7469, 7471, 7503, 7510, 7527, 7543, 7481, 7498, 7515, 7518, 7545, 7551, 7560-7592 7593-7613 7614-7645 12:40278025_T_A 7646-7693 7694-7742 7743-7794 12:40278154_A_G 7795-7831 7832-7865 7866-7903 12:40278167_C_T 7904-7950 7951-7998 7999-8049 12:40283881_G_A 8050-8073 8074-8099 8100-8127 12:40283915_T_C 8128-8174 8175-8224 8225-8276 12:40283947_C_T 8277-8324 8325-8374 8375-8426 12:40283966_G_A 8427-8473 8474-8523 8524-8575 12:40283985_C_A 8576-8622 8623-8672 8673-8724 12:40283990_T_C 8725-8772 8773-8822 8823-8874 12:40284061_A_G 8875-8921 8922-8967 8968-9017 12:40284076_C_A 9018-9056 9057-9088 9089-9126 12:40284139_T_C 9127-9164 9165-9193 9194-9226 12:40287363_C_A 9227-9266 9267-9300 9301-9338 12:40287438_A_G 9339-9385 9386-9435 9436-9487 12:40287461_A_G 9488-9535 9536-9581 9582-9631 12:40293555_CTC_CCT 9632-9671 9672-9711 9712-9753 12:40293607_C_T 9754-9798 9799-9842 9843-9888 12:40293613_G_A 9889-9936 9937-9986 9987-10038 12:40293624_G_C 10039-10086 10087-10136 10137-10188 12:40293626_G_A 10060, 10072, 10109, 10119, 10150, 10170, 10074, 10080, 10124, 10130, 10175, 10180, 10083-10084, 10133, 10136, 10185, 10188, 10189-10230 10231-10274 10275-10320 12:40293644_A_G 10321-10368 10369-10418 10419-10470 12:40294854_T_G 10471-10513 10514-10552 10553-10596 12:40294862_T_C 10597-10639 10640-10679 10680-10724 12:40294866_G_T 10622, 10628, 10644, 10747- 10681, 10758- 10725-10746 10757 10774 12:40294878_C_T 10775-10789 10790-10794 10795-10802 12:40295448_A_G 10803-10849 10850-10899 10900-10951 12:40295463_A_G 10952-10998 10999-11048 11049-11100 12:40295466_G_A 10955, 10965, 11002, 11016, 11067, 11076, 10991, 11101- 11025, 11041, 11091, 11095, 11144 11145-11190 11191-11238 12:40295535_A_T 11239-11277 11278-11315 11316-11359 12:40295566_A_G 11360-11407 11408-11457 11458-11509 12:40295574_A_C 11510-11557 11558-11607 11608-11659 12:40295581_G_T 11660-11706 11707-11745 11746-11791 12:40298330_A_G 11792-11836 11837-11880 11881-11929 12:40298333_C_T 11826, 11831, 11847, 11864, 11885, 11894, 11833, 11930- 11876, 11957- 11911, 11924, 11956 11972 11973-11994 12:40298346_G_A 11995-12042 12043-12088 12089-12138 12:40298433_C_G 12139-12186 12187-12236 12237-12288 12:40298479_G_T 12289-12335 12336-12382 12383-12434 12:40298488_A_G 12435-12481 12482-12530 12531-12582 12:40299125_A_G 12583-12622 12623-12659 12660-12703 12:40299190_C_T 12704-12751 12752-12800 12801-12852 12:40299196_A_C 12853-12900 12901-12950 12951-13002 12:40299212_G_A 13003-13049 13050-13099 13100-13151 12:40299255_T_C 13152-13199 13200-13249 13250-13300 12:40302866_A_G 13301-13347 13348-13384 13385-13427 12:40303952_C_A 13428-13471 13472-13517 13518-13566 12:40303972_C_G 13567-13613 13614-13663 13664-13715 12:40304000_G_A 13716-13762 13763-13811 13812-13863 12:40304004_A_G 13758, 13760, 13774, 13787, 13836, 13851, 13864-13909 13910-13956 13957-14006 12:40304035_A_C 14007-14049 14050-14099 14100-14151 12:40304040_G_C 14152-14193 14194-14227 14228-14272 12:40304040_G_T 14152-14153, 14194, 14202- 14235, 14240- 14157, 14162, 14203, 14208, 14241, 14248, 14166, 14169, 14215, 14219- 14257, 14262, 14182, 14185, 14220, 14226, 14264, 14270, 14273-14291 14292-14308 14272, 14309- 14331 12:40304078_G_A 14332-14352 14353-14370 14371-14392 12:40304141_C_T 14393-14430 14431-14455 14456-14488 12:40305776_C_T 14489-14505 14506-14522 14523-14541 12:40305781_T_A 14542-14568 14569-14594 14595-14622 12:40305791_C_G 14623-14666 14667-14711 14712-14758 12:40305871_G_A 14759-14790 14791-14816 14817-14849 12:40305942_G_A 14850-14868 14869-14882 14883-14900 12:40305946_T_A 14901-14930 14873, 14931- 14883, 14964- 14963 14998 12:40308467_G_T 14999-15018 15019-15034 15035-15053 12:40308476_A_G 15054-15100 15101-15145 15146-15193 12:40308481_G_A 15194-15230 15231-15266 15267-15309 12:40308543_GG_G 15310-15355 15356-15403 15404-15453 12:40308632_C_A 15454-15501 15502-15550 15551-15602 12:40308672_G_A 15603-15649 15650-15699 15700-15751 12:40309174_G_A 15752-15798 15799-15848 15849-15900 12:40309206_C_T 15901-15946 15947-15996 15997-16048 12:40309225_A_C 16049-16094 16095-16134 16135-16182 12:40309226_A_G 16058, 16060, 16102, 16117- 16145-16146, 16066, 16075, 16119, 16127, 16162-16163, 16077, 16084, 16129, 16223- 16174-16176, 16088, 16183- 16256 16257-16298 16222 12:40309230_A_G 16199, 16217, 16231, 16237, 16260, 16284, 16299-16343 16344-16382 16383-16429 12:40309236_G_A 16430-16445 16446-16453 16454-16465 12:40310434_C_A 16466-16510 16511-16560 16561-16612 12:40310434_C_G 16469, 16476- 16514, 16518, 16564, 16568, 16477, 16500, 16522, 16537, 16572, 16587- 16502, 16506, 16545, 16548, 16588, 16596- 16509-16510, 16550, 16557, 16597, 16600, 16613-16649 16559-16560, 16611-16612, 16650-16689 16690-16731 12:40310434_C_T 16469, 16476- 16514, 16518, 16564, 16568, 16477, 16500, 16522, 16537, 16572, 16587- 16502, 16506, 16545, 16548, 16588, 16596- 16509-16510, 16550, 16557, 16597, 16600, 16732-16768 16559-16560, 16611-16612, 16769-16808 16809-16850 12:40310435_G_A 16469, 16476, 16514, 16518, 16564, 16568, 16502, 16506, 16522, 16537, 16587-16588, 16509-16510, 16548, 16557, 16597, 16600, 16851-16889 16559-16560, 16611-16612, 16890-16931 16932-16975 12:40310436_C_T 16469, 16476, 16514, 16518, 16568, 16587- 16506, 16509, 16537, 16557, 16588, 16600, 16856, 16882, 16559-16560, 16611-16612, 16976-17015 16913, 16929, 16949, 16974, 17016-17057 17058-17101 12:40310437_G_C 16469, 16506, 16518, 16537, 16568, 16588, 16856, 16882, 16557, 16559, 16611-16612, 16999, 17013, 16913, 16929, 16949, 16974, 17102-17141 17033, 17049, 17063, 17099, 17142-17183 17184-17227 12:40310440_T_G 16999, 17013, 17033, 17049, 17063, 17099, 17119, 17134, 17147, 17181, 17184, 17219, 17228-17271 17272-17317 17318-17365 12:40310450_C_T 17366-17413 17414-17463 17464-17515 12:40310461_G_A 17516-17561 17562-17611 17612-17663 12:40310471_A_G 17664-17707 17708-17757 17758-17809 12:40310476_GAT_G 17810-17856 17857-17905 17906-17956 12:40310515_A_G 17957-18003 18004-18053 18054-18105 12:40310530_C_T 18106-18153 18154-18203 18204-18255 12:40310546_G_A 18256-18303 18304-18353 18354-18405 12:40310554_G_A 18406-18453 18454-18503 18504-18555 12:40310561_G_A 18556-18602 18603-18652 18653-18704 12:40314043_G_A 18705-18730 18731-18745 18746-18762 12:40314059_C_T 18763-18799 18800-18834 18835-18876 12:40314062_A_G 18783-18784, 18813, 18819, 18849, 18852, 18788, 18796, 18831, 18833, 18875-18876, 18877-18918 18919-18960 18961-19006 12:40314101_C_A 19007-19029 19030-19049 19050-19077 12:40315266_T_A 19078-19123 19124-19173 19174-19225 12:40319979_A_T 19226-19262 19263-19289 19290-19323 12:40319998_T_C 19324-19369 19370-19418 19419-19470 12:40320043_G_A 19471-19515 19516-19557 19558-19604 12:40320054_G_C 19605-19634 19635-19658 19659-19686 12:40320090_A_G 19687-19726 19727-19765 19766-19808 12:40320094_A_G 19712, 19809- 19739, 19853- 19793, 19897- 19852 19896 19942 12:40320097_T_C 19811, 19816, 19867, 19888, 19920, 19934, 19825, 19841, 19986-20030 20031-20077 19943-19985 12:40320119_A_G 20078-20123 20124-20167 20168-20215 12:40321114_A_G 20216-20259 20260-20303 20304-20353 12:40321123_T_C 20354-20396 20397-20438 20439-20482 12:40322037_C_T 20483-20526 20527-20566 20567-20612 12:40322038_G_A 20484-20485, 20528, 20533, 20570, 20573, 20490, 20492, 20535, 20550, 20579, 20596, 20509, 20518, 20650-20687 20688-20731 20520, 20613- 20649 12:40322047_G_A 20732-20774 20775-20824 20825-20876 12:40322047_G_T 20741, 20746, 20784, 20789, 20839, 20841, 20748, 20762, 20791, 20793- 20844, 20847, 20774, 20877- 20794, 20803, 20852, 20855, 20914 20809-20810, 20861-20862, 20813, 20819, 20865, 20871, 20915-20954 20955-20996 12:40322134_A_G 20997-21043 21044-21090 21091-21139 12:40322137_A_C 21006, 21030, 21074, 21078, 21120-21121, 21043, 21140- 21181-21223 21224-21268 21180 12:40322364_T_C 21269-21314 21315-21364 21365-21416 12:40322386_G_T 21417-21463 21464-21511 21512-21563 12:40322468_C_A 21564-21579 21580 21581-21583 12:40323244_C_G 21584-21623 21624-21661 21662-21701 12:40323255_A_G 21702-21729 21730-21752 21753-21779 12:40323256_T_C 21708, 21712, 21743-21744, 21768, 21773, 21716, 21720- 21746, 21749, 21776-21778, 21721, 21723, 21751, 21801- 21818-21838 21726, 21780- 21817 21800 12:40323260_G_T 21797, 21839- 21810, 21847- 21832, 21851- 21846 21850 21857 12:40323270_G_T 21858-21870 21871-21878 21879-21890 12:40323297_T_C 21891-21938 21939-21985 21986-22035 12:40328416_G_C 22036-22071 22072-22105 22106-22141 12:40335008_A_T 22142-22189 22190-22239 22240-22291 12:40335031_G_A 22292-22338 22339-22388 22389-22440 12:40335036_C_T 22441-22488 22489-22538 22539-22590 12:40335037_G_A 22447, 22463, 22495, 22514, 22545, 22567, 22466, 22470- 22516, 22519- 22571, 22573, 22471, 22475, 22520, 22524, 22576, 22585, 22484-22485, 22533, 22537, 22589-22590, 22591-22630 22631-22672 22673-22716 12:40335079_G_T 22717-22764 22765-22814 22815-22866 12:40340349_C_G 22867-22914 22915-22963 22964-23015 12:40340361_T_C 23016-23061 23062-23111 23112-23163 12:40340380_T_C 23164-23209 23210-23259 23260-23310 12:40340400_G_A 23311-23354 23355-23403 23404-23455 12:40340404_T_C 23352-23353, 23368-23369, 23426-23427, 23456-23499 23500-23547 23548-23597 12:40340432_A_G 23598-23644 23645-23694 23695-23746 12:40340436_A_T 23599, 23628, 23672, 23677, 23699, 23707, 23747-23792 23793-23840 23841-23890 12:40346818_T_C 23891-23932 23933-23966 23967-24008 12:40346826_ACTACT_A 24009-24028 24029-24036 24037-24048 12:40346884_A_G 24049-24091 24092-24133 24134-24177 12:40348402_G_GTGT 24178-24191 , 24192-24193 12:40348475_A_G 24194-24232 24233-24262 24263-24294 12:40348484_C_T 24295-24322 24323-24346 24347-24376 12:40351537_A_G 24377-24418 24419-24459 24460-24505 12:40351572_T_A 24506-24551 24552-24597 24598-24645 12:40351579_C_T 24646-24691 24692-24737 24738-24786 12:40351585_G_A 24787-24820 24821-24854 24855-24890 12:40351605_G_C 24891-24911 24912-24924 24925-24941 12:40351623_G_T 24942-24986 24987-25028 25029-25076 12:40351667_C_A 25077-25124 25125-25174 25175-25226 12:40351680_G_C 25227-25273 25274-25323 25324-25375 12:40351720_G_C 25376-25419 25420-25454 25455-25497 12:40351723_A_G 25408, 25417, 25429, 25454, 25467, 25469, 25419, 25498- 25542-25586 25587-25634 25541 12:40354304_T_C 25635-25678 25679-25720 25721-25766 12:40354305_G_T 25636, 25642, 25685, 25796- 25722, 25740, 25659, 25675, 25821 25748, 25822- 25677, 25767- 25853 25795 12:40354410_G_A 25854-25901 25902-25951 25952-26003 12:40356126_A_T 26004-26011 26012-26019 26020-26029 12:40356145_C_T 26030-26073 26074-26117 26118-26163 12:40359345_C_T 26164-26194 26195-26222 26223-26258 12:40359422_A_G 26259-26305 26306-26353 26354-26404 12:40363435_C_T 26405-26451 26452-26491 26492-26539 12:40363440_C_T 26540-26586 26587-26632 26633-26684 12:40363541_G_A 26685-26727 26728-26769 26770-26815 12:40364843_G_A 26816-26856 26857-26892 26893-26936 12:40364845_G_GGT 26937-26975 26976-27013 26904, 27014- 27055 12:40364884_G_A 27056-27080 27081-27097 27098-27122 12:40364927_A_T 27123-27169 27170-27219 27220-27271 12:40364960_A_G 27272-27318 27319-27368 27369-27420 12:40365044_C_T 27421-27468 27469-27518 27519-27570 12:40367012_T_A 27571-27614 27615-27646 27647-27685 12:40367050_A_G 27686-27733 27734-27783 27784-27835 12:40367648_AC_A 27836-27863 27864-27887 27888-27913 12:40367656_G_T 27914-27954 27955-27990 27991-28030 12:40367754_T_C 28031-28077 28078-28127 28128-28179 12:40367849_G_A 28180-28216 28217-28244 28245-28278 12:40367888_A_C 28279-28304 28305-28328 28329-28354 12:40367988_C_T 28355-28364 , , 12:40368042_A_G 28365-28394 28395-28417 28418-28444 12:40368073_A_C 28445-28476 28477-28491 28492-28514 12:40368170_C_T 28515-28547 28548-28566 28567-28593 12:40368262_C_T , 28594 28595-28596 12:40368380_T_A 28597-28602 28603 28604-28605 12:40368486_G_T 28606-28615 28616-28620 28621-28627 12:40368516_C_A 28628-28661 28662-28690 28691-28724 12:40368534_T_G 28725-28745 28746-28760 28761-28782 12:40368673_A_C 28783-28826 28827-28871 28872-28918 12:40368736_C_A 28919-28956 28957-28988 28989-29024 12:40368783_G_T 29025-29062 29063-29096 29097-29136 12:40368800_T_C 29137-29169 29170-29203 29204-29241 12:40368807_A_G 29242-29276 29277-29311 29312-29350 12:40368834_A_G 29351-29368 29369-29386 29387-29407 12:40368860_C_T 29408-29447 29448-29481 29482-29517 12:40368905_A_T 29518-29551 29552-29583 29584-29619 12:40368930_C_T 29620-29652 29653-29676 29677-29708 12:40368947_C_T 29709-29753 29754-29795 29796-29842 12:40368951_C_T 29752, 29843- 29771, 29876- 29804, 29909- 29875 29908 29945 12:40369044_T_C 29946-29948 29949 29950-29951 12:40369153_T_C 29952-29974 29975-29988 29989-30006 12:40369167_G_T 30007-30041 30042-30070 30071-30103 12:40222364_G_A 30104-30107 30108 30109-30111 rs1388598_REF 12:40222364_G_A 30107 , , rs1388598_SNP 12:40222612_A_G 30112-30145 30146-30171 30172-30203 rs28365207_REF 12:40222612_A_G 30128, 30136- 30156, 30164- 30184, 30194, rs28365207_SNP 30138, 30145, 30165, 30170- 30201-30203, 30204-30236 30171, 30237- 30270-30304 30269 12:40223400_T_C 30305 , 30306 rs7294619_REF 12:40223400_T_C 30305, 30307- 30312 30306, 30313- rs7294619_SNP 30311 30315 12:40223890_G_C 30316-30363 30364-30413 30414-30465 rs10878226_REF 12:40223890_G_C 30320, 30330, 30364, 30369, 30414, 30419, rs10878226_SNP 30337, 30343- 30374, 30387, 30424, 30437, 30344, 30349, 30393-30394, 30441, 30445- 30352, 30357- 30399, 30407- 30446, 30459- 30358, 30360, 30408, 30410, 30460, 30462, 30466-30503 30504-30543 30544-30585 12:40226614_A_T 30586-30627 30628-30659 30660-30697 rs1491943_REF 12:40226614_A_T 30590, 30593, 30633, 30637, 30666, 30671, rs1491943_SNP 30601, 30613, 30639, 30650, 30673-30674, 30616, 30698- 30653, 30735- 30688, 30762- 30734 30761 30794 12:40227006_C_G 30795-30841 30842-30891 30892-30943 rs1491942_REF 12:40227006_C_G 30798, 30802, 30845, 30848, 30893, 30896, rs1491942_SNP 30804, 30815, 30850, 30852, 30899, 30903, 30823, 30825, 30859, 30865, 30910, 30916, 30827, 30829, 30873, 30879, 30924, 30930, 30831, 30838, 30881, 30888, 30932, 30935, 30944-30981 30982-31021 31022-31063 12:40227013_G_A 30796-30797, 30843-30844, 30892-30895, rs1491941_REF 30800-30801, 30846-30849, 30897-30900, 30803-30814, 30851-30858, 30902-30909, 30816-30820, 30860-30861, 30911-30912, 30823-30825, 30863-30864, 30914-30915, 30828, 30830- 30866-30870, 30917-30921, 30835, 30838- 30873-30875, 30924-30926, 30840, 31064- 30878, 30880- 30929, 30931- 31077 30885, 30888- 30934, 30936- 30890, 31078- 30937, 30940- 31091 30942, 31092- 31105 12:40227013_G_A 30801, 30806, 30846, 30849, 30892, 30897, rs1491941_SNP 30816, 30819, 30854, 30866, 30905, 30917, 30824, 31067, 30869, 31081, 30920, 31093, 31074-31077, 31083, 31089- 31095, 31097, 31106-31143 31091, 31144- 31104-31105, 31183 31184-31225 12:40228090_A_G 31226-31247 31248-31269 31270-31291 rs11175645_REF 12:40228090_A_G 31226-31227, 31248-31249, 31271, 31274, rs11175645_SNP 31229, 31237- 31252, 31259- 31276, 31281- 31238, 31292- 31260, 31308- 31282, 31325- 31307 31324 31341 12:40228102_C_T 31226-31232, 31248-31256, 31270-31291, rs4567538_REF 31234, 31236- 31258-31269, 31388-31410 31247, 31342- 31365-31387 31364 12:40228102_C_T 31247, 31347, 31370, 31374, 31397, 31406, rs4567538_SNP 31351, 31360- 31384, 31387, 31409-31410, 31361, 31364, 31441-31473 31474-31510 31411-31440 12:40228988_C_T 31511-31558 31559-31604 31605-31654 rs12230685_REF 12:40228988_C_T 31511-31513, 31559-31562, 31605-31608, rs12230685_SNP 31515, 31521, 31569, 31574, 31610, 31621, 31526, 31537, 31585, 31599, 31623, 31648, 31544, 31552- 31689-31721 31722-31757 31553, 31655- 31688 12:40229925_G_A 31758-31804 31805-31854 31855-31906 rs11175655_REF 12:40229925_G_A 31763, 31767, 31810, 31813, 31860, 31863, rs11175655_SNP 31779, 31782, 31827, 31830, 31868, 31881, 31788, 31792, 31836, 31847, 31886, 31888, 31798, 31801, 31850, 31852- 31902, 31904- 31803-31804, 31854, 31945- 31906, 31985- 31907-31944 31984 32026 12:40229953_G_A 32027-32048 32049-32070 32071-32092 rs11175656_REF 12:40229953_G_A 32032-32033, 32054-32055, 32076-32078, rs11175656_SNP 32036-32037, 32059-32060, 32082-32083, 32039, 32093- 32062, 32110- 32127-32143 32109 32126 12:40229955_CTTTTTTTTTTT 32027-32031, 32049-32053, 32071-32075, TT_C 32033-32036, 32055-32058, 32077-32081, rs1402299410_REF 32038-32041, 32060-32063, 32083-32085, 32043, 32045- 32065, 32067- 32087, 32089- 32048 32070 32092 12:40229955_CTTTTTTTTTTT 32027-32031, 32049-32053, 32071-32075, C 32033-32036, 32055-32058, 32077-32081, rs780970090_REF 32038-32041, 32060-32063, 32083-32085, 32043, 32045- 32065, 32067- 32087, 32089- 32048 32070 32092 12:40229955_CT_C 32027-32031, 32049-32053, 32071-32075, rs1479481931_REF 32033-32036, 32055-32058, 32077-32081, 32038-32041, 32060-32063, 32083-32085, 32043, 32045- 32065, 32067- 32087, 32089- 32048 32070 32092 12:40229955_C_CT 32027-32031, 32049-32053, 32071-32075, rs71078229_REF 32033-32036, 32055-32058, 32077-32081, 32038-32041, 32060-32063, 32083-32085, 32043, 32045- 32065, 32067- 32087, 32089- 32048 32070 32092 12:40230508_A_G 32144-32190 32191-32238 32239-32290 rs11175658_REF 12:40230508_A_G 32144, 32148, 32191, 32195, 32239, 32243, rs11175658_SNP 32150, 32159, 32197, 32204, 32252, 32255, 32163, 32165, 32207, 32211, 32261, 32264, 32170, 32172, 32215, 32219, 32268, 32282- 32183, 32187, 32233, 32237, 32283, 32287, 32291-32328 32329-32368 32369-32410 12:40230668_CT_C 32411-32422 32423-32430 32431-32440 rs3057613_REF 12:40230668_C_CT 32411-32422 32423-32430 32431-32440 rs35642658_REF 12:40231279_G_A 32441-32463 32464-32488 32489-32515 rs74324737_REF 12:40231279_G_A 32441-32442, 32464, 32469, 32494, 32503- rs74324737_SNP 32453, 32458, 32478, 32483, 32504, 32510, 32463, 32516- 32488, 32540- 32515, 32566- 32539 32565 32593 12:40231365_C_A 32594-32598 32599-32605 32606-32616 rs2708441_REF 12:40231365_C_A 32594-32595, 32599-32600, 32606, 32609, rs2708441_SNP 32598, 32617- 32604-32605, 32613-32614, 32633 32634-32656 32616, 32657- 32682 12:40231390_C_CAAAA 32683-32697 32698-32713 32610, 32616, rs10631840_REF 32714-32728 12:40231390_C_CAAA 32683-32697 32698-32713 32610, 32616, rs35991926_REF 32714-32728 12:40231570_C_T 32729-32733 32734-32737 32738-32742 rs61931634_SNP 12:40234631_C_T 32743-32764 32765-32784 32785-32812 rs11175666_REF 12:40234631_C_T 32743, 32753, 32765, 32775, 32785, 32797, 32755, 32761, 32777, 32779, 32801, 32808, rs11175666_SNP 32813-32834 32835-32858 32859-32885 12:40235793_GTTT_G 32886-32888 32889-32890 32891-32893 rs751672176_REF 12:40235793_G_GTT 32886-32888 32889-32890 32891-32893 rs36024911_REF 12:40235793_G_GT 32886-32888 32889-32890 32891-32893 rs36024911_REF 12:40235794_T_TG 32886, 32888 32889 32892-32893 rs377568728_REF 12:40235794_T_TG 32888 , 32892 rs377568728_SNP 12:40235813_G_T 32894-32909 32910-32925 32926-32943 rs1279923177_REF 12:40235813_G_T 32894, 32896- 32910, 32912- 32926-32927, rs1279923177_SNP 32897, 32902, 32914, 32920, 32930-32931, 32904, 32944- 32953-32961 32937, 32962- 32952 32972 12:40237556_T_C 32973-33020 33021-33070 33071-33122 rs2249281_REF 12:40237556_T_C 32980, 32987, 33028, 33035, 33078, 33087- rs2249281_SNP 32989, 32997, 33037, 33040, 33088, 33091, 33006, 33008, 33046, 33055, 33093, 33098, 33011-33013, 33060-33062, 33107, 33112- 33016, 33123- 33066, 33161- 33113, 33118, 33160 33200 33201-33242 12:40237806_G_A 33243-33286 33287-33331 33332-33378 rs10878244_REF 12:40237806_G_A 33252, 33264, 33296, 33308- 33341-33342, rs10878244_SNP 33268, 33272, 33309, 33313, 33354-33355, 33282, 33286, 33331, 33416- 33359, 33451- 33379-33415 33450 33491 12:40237823_CAACA_C 33253, 33257, 33297, 33301- 33337, 33343, rs138000936_REF 33261, 33269- 33302, 33305, 33347-33348, 33270, 33272, 33314-33315, 33351, 33360- 33278, 33280- 33317, 33323, 33361, 33363, 33281, 33285, 33325-33326, 33369, 33371- 33492-33494 33330, 33495 33372, 33375, 33377, 33496 12:40237823_CAACA_C 33497 , rs138000936_SNP 12:40237989_T_C 33498-33540 33541-33576 33577-33617 rs10878245_REF 12:40237989_T_C 33499, 33501, 33543, 33547- 33578, 33580, rs10878245_SNP 33506-33507, 33548, 33563, 33585-33586, 33526-33527, 33565, 33656- 33600, 33606, 33529, 33618- 33695 33696-33737 33655 12:40238297_T_G 33738-33785 33786-33829 33830-33877 rs10878246_REF 12:40238297_T_G 33742, 33749, 33789, 33794, 33833, 33838, rs10878246_SNP 33759-33761, 33798, 33801, 33842, 33845, 33768, 33776, 33807-33808, 33850, 33853, 33781, 33784- 33815, 33823, 33860, 33864, 33785, 33878- 33827, 33829, 33869, 33874, 33915 33916-33955 33956-33997 12:40238298_C_T 33738-33754, 33786-33800, 33830-33845, rs10878247_REF 33756-33780, 33803-33829, 33847-33849, 33782-33785, 34000-34001 33851-33877, 33998-33999 34002-34003 12:40238298_C_T 33742, 33749, 33789, 33794, 33833, 33838, rs10878247_SNP 33760-33761, 33798, 33808, 33842, 33845, 33768, 33776, 33815, 33823, 33860, 33864, 33780, 33784- 33827-33829, 33869, 33874, 33785, 33998, 34000, 34032- 33877, 34002, 34004-34031 34053 34054-34081 12:40239156_G_T 34082-34129 34130-34179 34180-34231 rs954884_REF 12:40239156_G_T 34092, 34103, 34138, 34141, 34188, 34191, rs954884_SNP 34105, 34107, 34152, 34154, 34202, 34206, 34111, 34116, 34156, 34160, 34210-34211, 34121, 34123, 34169, 34171, 34215, 34221, 34126, 34129, 34176, 34179, 34223, 34231, 34232-34269 34270-34309 34310-34351 12:40239512_T_C 34352-34399 34400-34449 34450-34501 rs10878249_REF 12:40239512_T_C 34352, 34355, 34400, 34418, 34458, 34469, rs10878249_SNP 34370, 34378, 34426, 34429, 34477, 34480, 34381-34383, 34431, 34438, 34482, 34489, 34394-34395, 34443-34444, 34494, 34498- 34398, 34502- 34447-34448, 34499, 34501, 34539 34540-34579 34580-34621 12:40239686_G_A 34622-34652 34653-34682 34683-34716 rs10784444_REF 12:40239686_G_A 34634, 34641, 34657, 34664, 34688, 34695, rs10784444_SNP 34647, 34650, 34671, 34677, 34698, 34711, 34717-34739 34680, 34740- 34714, 34757- 34756 34775 12:40240356_T_C 34776-34787 34788-34799 34800-34813 rs6581622_REF 12:40240356_T_C 34776, 34778- 34788, 34790, 34803, 34806, rs6581622_SNP 34779, 34782, 34793, 34795, 34808, 34811, 34784, 34814- 34798, 34825- 34813, 34834- 34824 34833 34844 12:40241554_G_A 34845-34882 34883-34914 34915-34952 rs17519573_REF 12:40241554_G_A 34853, 34863, 34888, 34896, 34922-34923, rs17519573_SNP 34868, 34872, 34903, 34909, 34932, 34940, 34880, 34953- 34912, 34976- 34946, 34995- 34975 34994 35015 12:40242804_C_CAAAAAAA 35016-35028 35029-35041 35042-35056 AAAAAAAAAA rs35987733_REF 12:40242804_C_CAAAAAA 35016-35028 35029-35041 35042-35056 rs35987733_REF 12:40242818_G_A 35057-35062 35063-35068 35069-35076 rs868476079_SNP 12:40242819_G_A 35057, 35060- 35063-35064, 35070-35072, rs1399756142_SNP 35062, 35077- 35067-35068, 35075, 35085- 35080 35081-35084 35090 12:40242820_T_A_._SNP 35057, 35061- 35063-35064, 35070, 35072, 35062, 35077, 35067, 35082, 35075, 35086, 35091-35100 35101-35107 35108-35116 12:40242823_G_A 35077, 35096, 35082, 35131- 35086, 35135- rs1474763022_SNP 35117-35130 35134 35141 12:40242824_C_A 35096, 35119, 35155-35158 35159-35167 rs1450276224_SNP 35127, 35130, 35142-35154 12:40242827_T_A 35130, 35145, 35194-35212 35160, 35213- rs1191097078_SNP 35168-35193 35233 12:40242829_T_A 35179-35180, 35200-35201, 35221, 35229, rs_1425272990_SNP 35191, 35234- 35211, 35263- 35232, 35281- 35262 35280 35301 12:40242832_C_A 35254, 35260, 35266, 35277, 35283, 35287, rs1477924701_SNP 35302-35319 35320-35329 35330-35345 12:40245759_A_G 35346-35350 , 35351 rs2404580_REF 12:40245759_A_G 35348, 35352- 35377-35382 35351, 35383- rs2404580_SNP 35376 35392 12:40246099_C_T 35393-35404 35405-35412 35413-35422 rs2404581_REF 12:40246099_C_T 35395, 35398, 35405, 35407- 35414-35415, rs2404581_SNP 35401-35402, 35408, 35411- 35417-35418, 35404, 35423 35412, 35424 35421, 35425- 35427 12:40246358_T_C , , 35428 rs1388599_REF 12:40246358_T_C 35429-35438 , 35428 rs1388599_SNP 12:40246824_A_G 35439-35485 35486-35535 35536-35587 rs1388596_REF 12:40246824_A_G 35446, 35452, 35490, 35494, 35540, 35550, rs1388596_SNP 35456, 35459, 35504, 35507, 35558-35559, 35463, 35466- 35512, 35515- 35564, 35567- 35467, 35472- 35516, 35521, 35568, 35573, 35473, 35479, 35528, 35530, 35580, 35582, 35588-35625 35626-35665 35666-35707 12:40247890_A_G 35708-35747 35748-35785 35786-35825 rs10878258_REF 12:40247890_A_G 35715, 35718, 35749, 35756, 35786, 35788, rs10878258_SNP 35735, 35742, 35759, 35775, 35795, 35798, 35745, 35826- 35783, 35864- 35814, 35901- 35863 35900 35939 12:40250503_T_C 35940-35943 35944-35946 35947-35950 rs12371502_REF 12:40250503_T_C 35940-35943 35944-35946 35947-35951 rs12371502_SNP 12:40250565_C_T 35952-35980 35981-36014 36015-36054 rs11175729_REF 12:40250565_C_T 35956-35957, 35986, 35992, 36022, 36024, rs11175729_SNP 35962, 35968, 35998, 36002, 36036, 36041- 35971, 35975, 36006, 36008, 36042, 36046, 36055-36075 36010, 36076- 36048, 36050, 36099 36100-36126 12:40250950_A_G 36127-36139 36140-36156 36157-36174 rs17443414_REF 12:40250950_A_G 36127-36128, 36140-36142, 36157-36159, rs17443414_SNP 36130, 36132, 36144, 36149, 36161, 36166, 36135, 36175- 36204-36236 36237-36273 36203 12:40251002_G_GT 36274-36291 36292-36309 36310-36327 rs1486599371_REF 12:40251072_C_T 36328-36349 36350-36367 36368-36387 rs7134379_REF 12:40251072_C_T 36330, 36332, 36353, 36356, 36371, 36375- rs7134379_SNP 36336, 36339, 36358, 36363, 36376, 36378, 36345, 36388- 36365, 36399- 36385, 36410- 36398 36409 36422 12:40251455_C_A 36423-36467 36468-36511 36512-36559 rs7955902_REF 12:40251455_C_A 36425, 36431- 36470, 36474, 36514, 36520, rs7955902_SNP 36433, 36443, 36476-36477, 36522-36523, 36446, 36457, 36490, 36501, 36537, 36554, 36560-36593 36594-36625 36626-36661 12:40251794_A_G 36662-36702 36703-36733 36734-36772 rs1491939_REF 12:40251794_A_G 36664, 36669, 36706, 36719, 36736, 36741, rs1491939_SNP 36677, 36684, 36721, 36811- 36755, 36757, 36686, 36688, 36848 36759, 36849- 36773-36810 36890 12:40251828_G_A 36891-36925 36926-36959 36960-36995 rs1491938_REF 12:40251828_G_A 36894, 36896, 36929, 36931, 36963, 36965- rs1491938_SNP 36903, 36914, 36939-36940, 36966, 36975, 36922, 36996- 36949, 37023- 36985, 37038- 37022 37037 37056 12:40253146_G_A 37057-37101 37102-37138 37139-37183 rs7969677_REF 12:40253146_G_A 37060, 37067, 37109, 37117, 37142, 37149, rs7969677_SNP 37073, 37075, 37132 37173, 37205- 37091, 37099, 37211 37101, 37184- 37204 12:40255521_C_T 37212-37256 37257-37301 37302-37349 rs2723265_REF 12:40255521_C_T 37220, 37231- 37267, 37277- 37311, 37313, rs2723265_SNP 37232, 37237, 37278, 37283, 37323-37324, 37242, 37244, 37289, 37291, 37329, 37336, 37246, 37350- 37383-37407 37339, 37408- 37382 37440 12:40255947_G_C 37441-37477 37478-37496 37497-37522 rs7132171_REF 12:40255947_G_C 37446, 37458- 37482, 37490, 37501, 37509, rs7132171_SNP 37459, 37466, 37556-37572 37513, 37573- 37523-37555 37595 12:40256086_G_A 37596-37626 37627-37655 37656-37687 rs10784451_REF 12:40256086_G_A 37601, 37608- 37629, 37638- 37657, 37660, 37639, 37649, 37670-37671, rs10784451_SNP 37609, 37617, 37711-37730 37681, 37731- 37620, 37688- 37752 37710 12:40256173_G_A 37753-37771 37772-37794 37795-37821 rs10784452_REF 12:40256173_G__A 37753, 37755- 37773, 37776, 37799, 37805- rs10784452_SNP 37756, 37762, 37782, 37793- 37806, 37819, 37771, 37822- 37794, 37846- 37821, 37874- 37845 37873 37903 12:40256549_C_T 37904-37935 37936-37955 37956-37983 rs1907633_REF 12:40256549_C_T 37907, 37912, 37936-37937, 37956-37957, rs1907633_SNP 37915-37916, 37939, 37941, 37960, 37963, 37919, 37922, 37943, 37946, 37969, 37971, 37924, 37928, 37948, 37950, 37974, 37976, 37933-37934, 37953-37954, 37981-37982, 37984-37988 37989-37993 37994-38001 12:40256647_T_C 38002-38008 38009-38010 38011-38013 rs1491936_REF 12:40256647_T_C 38002-38004, 38009-38010, 38011-38013, rs1491936_SNP 38007-38008, 38035-38039 38040-38048 38014-38034 12:40256700_G_A 38049-38096 38097-38146 38147-38198 rs1907632_REF 12:40256700_G_A 38050, 38053, 38097, 38099, 38147, 38149, rs1907632_SNP 38059, 38067, 38102, 38105, 38152, 38155, 38072, 38076, 38109, 38117, 38159, 38163, 38079-38081, 38129-38131, 38168, 38182, 38090, 38199- 38140, 38237- 38185, 38192, 38236 38276 38277-38318 12:40258592_A_G 38319-38366 38367-38416 38417-38468 rs2046928_REF 12:40258592_A_G 38324, 38326, 38372, 38374- 38424-38425, rs2046928_SNP 38342-38344, 38375, 38387, 38432, 38435, 38348, 38354, 38392-38393, 38439, 38444- 38360-38361, 38398, 38404, 38445, 38450, 38365, 38469- 38410-38411, 38462-38463, 38506 38507-38546 38547-38588 12:40258718_T_C 38589-38636 38637-38686 38687-38738 rs2723264_REF 12:40258718_T_C 38591, 38599- 38639, 38641, 38691, 38699, rs2723264_SNP 38600, 38605- 38649, 38654- 38704-38706, 38607, 38609, 38656, 38658, 38708, 38710, 38611, 38618, 38660, 38667, 38714, 38719, 38631, 38739- 38680, 38777- 38732, 38817- 38776 38816 38858 12:40259911_C_T 38859-38896 38897-38932 38933-38969 rs17465870_REF 12:40259911_C_T 38876, 38884, 38926, 39005- 38950, 38966, 38889, 38893, rs17465870_SNP 38970-39004 39039 39040-39074 12:40260186_C_T 39075-39121 39122-39164 39165-39209 rs10878278_REF 12:40260186_C_T 39075, 39078- 39122, 39125, 39193, 39196, 39079, 39085, 39150, 39248- 39089, 39104, rs10878278_SNP 39107, 39115, 39283 39204, 39284- 39117, 39210- 39325 39247 12:40261214_T_C 39326-39329 , , rs11175766_REF 12:40261214_T_C 39326-39335 , 39336 rs11175766_SNP 12:40261905_C_T 39337-39350 39351-39354 39355-39360 rs11564183_REF 12:40262377_A_G 39361-39398 39399-39435 39436-39475 rs11175769_REF 12:40262377_A_G 39373, 39382, 39411, 39419, 39449, 39453, rs11175769_SNP 39386, 39388, 39425, 39428, 39458, 39464, 39391, 39395, 39433-39434, 39467, 39473, 39397, 39476- 39509-39541 39475, 39542- 39508 39577 12:40262593_C_T 39578-39619 39620-39662 39663-39706 rs17465912_REF 12:40262593_C_T 39587, 39601, 39644, 39652, 39686, 39688, rs17465912_SNP 39609, 39614, 39657, 39660, 39700, 39703, 39617, 39707- 39744-39778 39706, 39779- 39743 39815 12:40262875_C_A 39816-39863 39864-39913 39914-39965 rs17443552_REF 12:40262875_C_A 39817-39818, 39865-39866, 39915, 39920, rs17443552_SNP 39824, 39828- 39872, 39876, 39923, 39927, 39829, 39837, 39885, 39888- 39936, 39939- 39840, 39850, 39889, 39899, 39940, 39953, 39854, 39863, 39902, 39904, 39955, 39962, 39966-40003 40004-40043 40044-40085 12:40263160_T_A 40086-40088 , rs17490817_REF 12:40263160_T_A 40086-40088 , rs17490817_SNP 12:40263401_G_GA 40089-40120 40121-40142 40143-40170 rs11430638_REF 12:40263401_G_GA 40094, 40104, 40122, 40129, 40146, 40154, rs11430638_SNP 40115, 40119, 40134, 40141, 40160, 40166, 40171-40192 40193-40204 40205-40222 12:40263423_G_A 40094, 40103, 40122, 40128, 40146, 40153, rs4293189_REF 40110, 40114, 40134-40136, 40159-40160, 40223-40265 40139, 40266- 40162-40163, 40309 40166-40167, 40310-40353 12:40263423_G_A 40103, 40114, 40128, 40135- 40153, 40162- rs4293189_SNP 40225, 40227, 40136, 40268, 40163, 40167, 40233, 40235, 40271-40272, 40312, 40315- 40238, 40240, 40280, 40283, 40316, 40324, 40242, 40246, 40285, 40291, 40333, 40336, 40354-40391 40392-40425 40426-40463 12:40263735_A_G 40464-40495 40496-40524 40525-40555 rs10784461_REF 12:40263735_A_G 40478, 40487- 40510, 40518, 40536, 40549, rs10784461_SNP 40488, 40491, 40520, 40523, 40551, 40554, 40494, 40556- 40591-40619 40620-40652 40590 12:40263898_C_G 6533, 6628, 6560, 6577, 6599, 6603, rs7308720_REF 6631, 6644, 6663, 6665, 6622, 6691, 6651, 6659, 6668, 6678, 6693, 6696, 6727, 6737- 6689, 6762, 6701, 6708, 6738, 6742, 6771-6772, 6804-6805, 6747, 40653- 6779, 6783, 6814-6815, 40685 40686-40716 6819, 40717- 40750 12:40263898_C_G 6628, 6631, 6663, 6665, 6691, 6693, rs7308720_SNP 6644, 40653, 6668, 40687, 6701, 40720, 40656, 40660, 40696, 40789- 40731, 40827- 40665, 40751- 40826 40868 40788 12:40263969_C_G 40869-40913 40914-40956 40957-41003 rs10784462_REF 12:40263969_C_G 40870, 40879, 40931, 40938, 40970, 40975, rs10784462_SNP 40887, 40889, 40953, 41042- 40977, 40984, 40895, 40905, 41081 40997, 41082- 40910, 41004- 41123 41041 12:40264403_G_A 41124-41127 41128-41133 41134-41139 rs10878284_REF 12:40265294_C_T 41140-41187 41188-41237 41238-41289 rs11175780_REF 12:40265294_C_T 41157, 41168, 41217, 41219, 41247, 41250, rs11175780_SNP 41170, 41174, 41223-41224, 41269, 41271, 41177, 41182- 41227, 41232- 41275-41276, 41183, 41185- 41233, 41235- 41285, 41287- 41187, 41290- 41237, 41328- 41289, 41368- 41327 41367 41409 12:40265732_C_T 41410-41450 41451-41493 41494-41537 rs11175784_REF 12:40265732_C_T 41411, 41414, 41451, 41453, 41494, 41496, rs11175784_SNP 41421, 41424, 41456, 41463, 41499, 41506, 41442, 41538- 41485, 41570- 41516, 41604- 41569 41603 41640 12:40266292_C_A 41641 , 41642 rs11175787_REF 12:40267078_A_T , 41643 41644-41646 rs144463374_SNP 12:40267251_A_T 41647-41691 41692-41738 41739-41787 rs73108329_REF 12:40267251_A_T 41655-41656, 41701, 41708, 41742, 41744, rs73108329_SNP 41663, 41668, 41710, 41714, 41750, 41757, 41679, 41682, 41726, 41729, 41759, 41763, 41685, 41688, 41735, 41826- 41784, 41866- 41788-41825 41865 41907 12:40267479_T_C 41908-41953 41954-42003 42004-42055 rs12426891_REF 12:40267479_T_C 41908, 41914, 41962, 41966- 42012, 42016- rs12426891_SNP 41916, 41920- 41967, 41973, 42017, 42026, 41921, 41927, 41976, 41984- 42035, 42041- 41930, 41938- 41985, 41991, 42042, 42047- 41939, 41948, 41996-41997, 42048, 42054, 42056-42093 42094-42133 42134-42175 12:40267601_A_T 42176-42223 42224-42273 42274-42325 rs4768224_REF 12:40267601_A_T 42184, 42191, 42232, 42235, 42282-42283, rs4768224_SNP 42201-42205, 42240, 42251, 42286, 42291, 42214-42216, 42253-42255, 42302-42303, 42326-42363 42264-42266, 42306-42307, 42364-42403 42317-42318, 42404-42445 12:40268237_G_A 42446-42493 42494-42543 42544-42595 rs11175793_REF 12:40268237_G_A 42450, 42459, 42498, 42507, 42548, 42557, rs11175793_SNP 42463-42464, 42511, 42514, 42564-42565, 42466, 42469, 42521, 42524, 42572, 42575, 42473, 42476, 42532, 42534, 42583, 42587, 42485, 42487, 42536, 42541, 42592, 42594, 42596-42633 42634-42673 42674-42715 12:40269794_G_T 42716-42763 42764-42813 42814-42865 rs10784470_REF 12:40269794_G_T 42717, 42721- 42764, 42766, 42814-42815, rs10784470_SNP 42722, 42728, 42770-42771, 42817, 42821- 42731, 42738- 42777, 42780- 42822, 42828, 42739, 42741, 42781, 42788, 42831-42832, 42743, 42755, 42793, 42805, 42844, 42850, 42866-42901 42902-42923 42924-42957 12:40270116_C_G 42958-42993 42994-43015 43016-43041 rs12146857_REF 12:40270116_C_G 42964, 42968, 42996, 42998, 43019, 43022, rs12146857_SNP 42976, 42984, 43001, 43003, 43025, 43029, 42987, 43042- 43010, 43073- 43035, 43090- 43072 43089 43110 12:40270841_T_A 43111-43131 43132-43145 43146-43163 rs11564207_REF 12:40270841_T_A 43119, 43131, 43183-43196 43197-43214 rs11564207_SNP 43164-43182 12:40271425_G_C 43215-43262 43263-43309 43310-43358 rs7308193_REF 12:40271425_G_C 43222, 43225, 43272, 43295, 43319, 43321, rs7308193_SNP 43231, 43246, 43302, 43304, 43341, 43351, 43249, 43251, 43306, 43308- 43355, 43357- 43256, 43258, 43309, 43397- 43358, 43437- 43260, 43262, 43436 43478 43359-43396 12:40272958_A_T 43479-43524 43525-43566 43567-43616 rs10506150_REF 12:40272958_A_T 43495, 43498- 43539, 43542- 43579, 43584, rs10506150_SNP 43499, 43501, 43543, 43545, 43587-43588, 43509, 43512, 43552-43553, 43593, 43599- 43517-43519, 43556, 43561, 43600, 43603, 43524, 43617- 43565-43566, 43610, 43615, 43652 43653-43684 43685-43724 12:40273002_GAC_G 43725-43772 43773-43822 43823-43874 rs76660489_REF 12:40273002_GAC_G 43727, 43732, 43775, 43781, 43825, 43827, rs76660489_SNP 43741, 43749- 43789, 43794, 43832, 43845, 43750, 43759, 43798-43799, 43849-43850, 43769, 43772, 43809, 43822, 43853, 43860, 43875-43913 43914-43954 43955-43997 12:40273073_C_G 43998-44028 44029-44044 44045-44065 rs10878299_REF 12:40273073_C_G 44007, 44011, 44035, 44093- 44055, 44061, rs10878299_SNP 44014, 44023, 44107 44108-44126 44066-44092 12:40275500_TTAAGAA_T 44127-44155 44156-44183 44184-44215 rs373619252_REF 12:40275500_TTAAGAA_T 44130-44131, 44157-44158, 44186, 44193, rs373619252_SNP 44139, 44142, 44164, 44166, 44195, 44198, 44145, 44216- 44172, 44236- 44204, 44254- 44235 44253 44275 12:40276024_T_A 44276-44277 , , rs10878302_REF 12:40276024_T_A 44276-44277 , , rs10878302_SNP 12:40276137_T_C 44278-44300 44301-44308 44309-44323 rs11564129_REF 12:40276137_T_C 44283, 44288- 44301, 44303, 44310, 44313- rs11564129_SNP 44289, 44291- 44306-44308, 44314, 44318, 44292, 44295, 44353-44373 44320-44321, 44324-44352 44374-44397 12:40276592_G_A 44398-44427 44428-44460 44461-44495 rs7971935_REF 12:40276592_G_A 44405, 44417, 44449, 44459- 44469, 44483, rs7971935_SNP 44425-44427, 44460, 44516- 44495, 44538- 44496-44515 44537 44561 12:40276874_C_T 44562-44570 44571-44583 44584-44600 rs11175836_REF 12:40276874_C_T 44563, 44566, 44571, 44576, 44584, 44590, rs11175836_SNP 44601-44628 44581, 44629- 44596, 44599, 44661 44662-44695 12:40276999_C_T 44696-44731 44732-44763 44764-44798 rs4272849_REF 12:40276999_C_T 44713, 44717, 44738, 44747, 44770, 44783, rs4272849_SNP 44728, 44799- 44750, 44825- 44792, 44851- 44824 44850 44878 12:40277068_T_C 44879-44889 44890-44903 44904-44923 rs10784485_REF 12:40277068_T_C 44879-44880, 44891-44892, 44906, 44908- rs10784485_SNP 44882, 44885- 44894-44896, 44910, 44916, 44886, 44924- 44900, 44945- 44918-44919, 44944 44968 44969-44995 12:40277196_C_A 44996-45033 45034-45063 45064-45099 rs10506151_REF 12:40277196_C_A 44996, 44998, 45034-45035, 45064-45065, rs10506151_SNP 45002, 45010, 45038, 45044, 45077, 45085, 45023, 45100- 45051, 45123- 45094, 45146- 45122 45145 45170 12:40279458_T_A 45171-45206 45207-45237 45238-45274 rs7309197_REF 12:40279458_T_A 45177, 45185, 45220, 45235, 45245, 45253, rs7309197_SNP 45191, 45200, 45306-45334 45335-45369 45204, 45275- 45305 12:40279566_G_A 45370-45395 45396-45403 45404-45417 rs7134572_REF 12:40279566_G_A 45373, 45382- 45396, 45398, 45404, 45407, rs7134572_SNP 45384, 45418- 45401-45402 45411, 45414- 45419 45415 12:40280237_G_C 45420-45422 45423-45427 45428-45434 rs11175843_REF 12:40280237_G_C 45421-45422, 45424, 45426- 45429, 45431- rs11175843_SNP 45435-45446 45427, 45447- 45432, 45434, 45461 45462-45483 12:40280621_T_TTAAAATAA 45484-45504 45505-45527 45528-45554 AA rs147801391_REF 12:40280621_T_TTAAAA 45484-45504 45505-45527 45528-45554 rs147801391_REF 12:40280621_T_TTAAATAAA 45484-45504 45505-45527 45528-45554 ATAAAA rs147801391_REF 12:40280621_T_TTAAATAAA 45484, 45490, 45505, 45511, 45528, 45533, ATAAAA 45495, 45555- 45517, 45519, 45536, 45542, rs147801391_SNP 45568 45569-45581 45545, 45582- 45595 12:40280621_T_TTAAATAAA 45484-45504 45505-45527 45528-45554 A rs147801391_REF 12:40280621_T_TTAAATAAA 45484, 45490, 45505, 45511, 45528, 45533, A 45495, 45555- 45517, 45519, 45536, 45542, rs147801391_SNP 45568 45569-45581 45545, 45582- 45595 12:40280622_TA_T 45484, 45486- 45505, 45507- 45528, 45530- rs376096297_REF 45504 45527 45554 12:40280622_TA_T 45484, 45490, 45505, 45511, 45528, 45533, rs376096297_SNP 45495, 45555- 45517, 45519, 45536, 45542, 45568 45569-45581 45545, 45582- 45595 12:40280792_T_C 45596-45622 45623-45651 45652-45682 rs7488873_REF 12:40280792_T_C 45612-45613, 45640, 45645, 45669-45670, rs7488873_SNP 45617, 45620, 45648-45649, 45676, 45680, 45622, 45683- 45651, 45704- 45682, 45719- 45703 45718 45738 12:40281250_A_G 45739-45786 45787-45834 45835-45886 rs11175845_REF 12:40281250_A_G 45745, 45753, 45792, 45800, 45841-45842, rs11175845_SNP 45755, 45758, 45802, 45805, 45850, 45852, 45761, 45765, 45808, 45812, 45855, 45862, 45768, 45772, 45815-45816, 45866-45867, 45775, 45780, 45824, 45829, 45875, 45880, 45887-45924 45925-45964 45965-46006 12:40281806_A_G 46007-46052 46053-46096 46097-46144 rs7979341_REF 12:40281806_A_G 46017, 46019, 46056, 46065, 46098, 46101, rs7979341_SNP 46027, 46029- 46073, 46087, 46111, 46121- 46030, 46032, 46096, 46183- 46122, 46135, 46043, 46052, 46222 46144, 46223- 46145-46182 46264 12:40282009_GCTTCCCTTCC 46265-46298 46299-46338 46339-46382 CTTCCCTTCCCTTCC_G rs1226327150_REF 12:40282053_CCCTTCCCTTC , 46383 46384-46390 CCTTCCCTTCCCTTCCCTTC CCTTCTCTT_C rs777452337_SNP 12:40282112_TCCCTC_T 46391-46401 46383, 46402- 46384, 46386- rs1339001348_REF 46415 46388, 46416- 46432 12:40282398_G_T 46433-46475 46476-46514 46515-46562 rs11175847_REF 12:40282398_G_T 46435, 46443, 46477, 46479, 46516, 46519, rs11175847_SNP 46445, 46447, 46486, 46488- 46524, 46529, 46450-46451, 46489, 46492, 46531, 46533- 46464, 46467, 46504, 46506, 46534, 46537, 46563-46589 46590-46612 46551, 46553, 46613-46639 12:40283044_TA_T 46640-46644 46645-46649 46650-46654 rs112859457_REF 12:40283045_A_T 46641-46644 46645-46646, 46650, 46652- rs17466080_REF 46648-46649 46654 12:40283045_A_T 46641-46644 46645-46646, 46650, 46652- rs17466080_SNP 46648-46649 46654 12:40283206_G_A 46655-46702 46703-46752 46753-46804 rs11175852_REF 12:40283206_G_A 46657, 46666, 46705, 46714- 46755, 46759, rs11175852_SNP 46668, 46671, 46715, 46717, 46765-46766, 46680, 46693- 46720, 46735, 46768, 46771, 46695, 46698- 46744-46745, 46785, 46787, 46699, 46805- 46748-46749, 46797, 46800, 46842 46843-46882 46883-46924 12:40283227_A_C 46657-46658, 46705-46706, 46755-46756, rs10784486_REF 46664, 46668, 46712, 46717, 46759, 46763, 46671, 46674, 46720-46721, 46768, 46771- 46925-46966 46723, 46735, 46772, 46774, 46967-47008 46778, 46787, 47009-47050 12:40283227_A_C 46658, 46664, 46706, 46712, 46756, 46763, rs10784486_SNP 46674, 46925- 46721, 46723, 46772, 46774, 46927, 46932, 46967-46968, 46778, 47009, 46939, 46949, 46974, 46981, 47016, 47022, 46955, 47051- 46983, 46997, 47024, 47026, 47088 47089-47128 47129-47170 12:40283782_G_T 47171-47217 47218-47248 47249-47288 rs73108368_REF 12:40283782_G_T 47172, 47175, 47232, 47241, 47255, 47279, rs73108368_SNP 47182, 47189, 47246, 47310- 47284, 47287, 47195, 47206, 47319 47320-47336 47212-47213, 47215, 47289- 47309 12:40284261_AT_A 47337-47354 47355-47364 47365-47376 rs398019201_REF 12:40284539_C_T 47377-47405 47406-47431 47432-47464 rs17519846_REF 12:40284539_C_T 47382, 47390- 47410-47411, 47437-47438, 47391, 47393- 47419, 47421, 47440, 47451, 47394, 47398, 47424, 47426, 47456-47457, rs17519846_SNP 47400, 47402, 47428, 47478 47459, 47461, 47465-47477 47479-47483 12:40285452_G_A 47484-47519 47520-47551 47552-47587 rs17466150_REF 12:40285452_G_A 47492-47493, 47525-47526, 47559-47561, rs17466150_SNP 47503, 47509, 47540-47541, 47576-47577, 47512, 47588- 47544, 47611- 47634-47658 47610 47633 12:40285467_T_A 47484-47485, 47520-47527, 47552-47553, rs11175862_REF 47489-47493, 47531, 47534- 47556-47562, 47500, 47503- 47537, 47539- 47567, 47570- 47506, 47508- 47541, 47544- 47573, 47575- 47509, 47512- 47546, 47550, 47578, 47580- 47514, 47518, 47673-47682 47582, 47586, 47659-47672 47683-47694 12:40285467_T_A 47504, 47506, 47527, 47537, 47562, 47573, rs11175862_SNP 47513-47514, 47545-47546, 47578, 47582, 47518, 47695- 47550, 47722- 47586, 47747- 47721 47746 47775 12:40286069_A_G 47776-47822 47823-47870 47871-47920 rs4310676_REF 12:40286069_A_G 47782, 47784, 47826, 47830, 47874, 47880, rs4310676_SNP 47788, 47809- 47832, 47836, 47884, 47895, 47810, 47812, 47858, 47860, 47909-47910, 47819-47820, 47868, 47870, 47918, 47920, 47822, 47921- 47959-47998 47999-48040 47958 12:40287115_T_C 48041-48082 48083-48115 48116-48157 rs11564150_REF 12:40287115_T_C 48049, 48060, 48089, 48094, 48128, 48132, rs11564150_SNP 48067, 48069, 48104, 48107, 48135, 48144, 48073, 48076, 48110, 48112, 48147, 48153, 48079, 48082, 48115, 48196- 48156-48157, 48158-48195 48235 48236-48277 12:40287139_A_G 48278-48325 48093-48094, 48127-48128, rs4318033_REF 48326-48373 48131-48132, 48374-48421 12:40287139_A_G 48278, 48283- 48093, 48326, 48127, 48131, rs4318033_SNP 48285, 48287, 48331-48333, 48374, 48378, 48298, 48302- 48335, 48342, 48381, 48384, 48303, 48309, 48350, 48357, 48391, 48398, 48312, 48422- 48360, 48460- 48405, 48408, 48459 48499 48500-48541 12:40287255_C_T 48542-48575 48576-48609 48610-48645 rs4473003_REF 12:40287255_C_T 48555, 48561, 48580, 48590, 48611, 48616, rs4473003_SNP 48569, 48572, 48595, 48603, 48631, 48639, 48574, 48646- 48608, 48671- 48644, 48690- 48670 48689 48712 12:40287340_C_CT 48713-48743 48744-48772 48773-48804 rs1429478461_REF 12:40287340_C_CT 48716, 48722, 48753, 48763- 48783-48784, rs1429478461_SNP 48731, 48734- 48764, 48770, 48791, 48795, 48735, 48741, 48772, 48823- 48801, 48804, 48805-48822 48838 48839-48858 12:40287550_T_TA 48859-48862 48863-48865 48866-48869 rs886049356_REF 12:40287684_A_G 48870-48881 48882-48890 48891-48905 rs11564270_REF 12:40287684_A_G 48870-48871, 48882-48883, 48892, 48895, rs11564270_SNP 48873-48875, 48886-48890, 48898-48899, 48877-48881, 48920-48927 48901, 48903, 48906-48919 48905, 48928- 48941 12:40288395_C_G 48942-48986 48987-49032 49033-49080 rs17519881_REF 12:40288395_C_G 48947, 48951, 48992, 48996, 49038, 49045, rs17519881_SNP 48959, 48963, 49008, 49012, 49055, 49059, 48967, 48970, 49014, 49022, 49061, 49063, 48976, 49081- 49119-49158 49159-49200 49118 12:40288788_G_T 49201-49203 , 49204 rs17466178_REF 12:40288788_G_T 49203 , , rs17466178_SNP 12:40290820_T_G 49205-49236 49237-49257 49258-49285 rs17466185_REF 12:40290820_T_G 49205-49206, 49239-49240, 49258, 49262, rs17466185_SNP 49210-49211, 49246-49247, 49269-49270, 49215, 49218, 49317-49345 49280, 49346- 49224, 49286- 49378 49316 12:40291078_A_C 49379-49383 49384-49390 49391-49398 rs11175902_SNP 12:40291483_C_T 49399-49444 49445-49480 49481-49522 rs76033741_REF 12:40291483_C_T 49400, 49403, 49446, 49453, 49481, 49484, rs76033741_SNP 49410, 49415, 49458, 49463, 49491, 49493, 49422-49423, 49471, 49473, 49497, 49512, 49430, 49434, 49477, 49479, 49519, 49521, 49439, 49444, 49544-49562 49563-49588 49523-49543 12:40292044_C_T 49589-49635 49636-49685 49686-49737 rs56325408_REF 12:40292044_C_T 49590, 49592, 49636, 49638, 49686-49687, rs56325408_SNP 49597, 49599, 49640, 49645, 49689, 49691, 49613, 49621, 49647, 49661, 49698, 49712, 49624-49625, 49672, 49674, 49725, 49729, 49629, 49633, 49678, 49685, 49736-49737, 49738-49775 49776-49815 49816-49857 12:40292236_G_T 49858-49902 49903-49946 49947-49996 rs7957754_REF 12:40292236_G_T 49862, 49872- 49913, 49916- 49958, 49963, rs7957754_SNP 49873, 49885, 49917, 49927, 49971, 49976- 49891-49892, 49933-49935, 49977, 49983- 49897-49899, 49939, 49942- 49984, 49989, 49997-50006 49943, 50007- 49992-49993, 50014 50015-50024 12:40292929_A_G 50025-50037 50038-50046 50047-50058 rs11175911_REF 12:40292929_A_G 50031, 50034, 50044, 50078- 50054, 50058, rs11175911_SNP 50036-50037, 50088 50089-50103 50059-50077 12:40293043_A_G 50104-50115 50116-50119 50120-50124 rs12369882_REF 12:40293043_A_G 50107, 50109- 50116-50119, 50120-50124, rs12369882_SNP 50110, 50114- 50136-50144 50145-50155 50115, 50125- 50135 12:40293233_C_T 50156-50202 50203-50252 50253-50304 rs11175913_REF 12:40293233_C_T 50161-50163, 50203, 50210, 50253-50254, rs11175913_SNP 50168, 50177, 50212-50213, 50261, 50263, 50185, 50190- 50218, 50227, 50269, 50278- 50191, 50197, 50235, 50240- 50279, 50292- 50200, 50305- 50241, 50247, 50293, 50299, 50342 50343-50382 50383-50424 12:40293852_A_G 50425-50456 50457-50473 50474-50497 rs4483664_REF 12:40293852_A_G 50436, 50438, 50462-50463, 50475, 50483- rs4483664_SNP 50442, 50444, 50467, 50469, 50484, 50488, 50446, 50453, 50472, 50536- 50495, 50568- 50498-50535 50567 50605 12:40293893_GCA_G 50606-50639 50640-50660 50661-50684 rs66487867_REF 12:40293893_GCA_G 50607, 50616, 50640, 50645, 50661, 50666, rs66487867_SNP 50618, 50620- 50647, 50649, 50668, 50670, 50621, 50685- 50658, 50695- 50682, 50699- 50694 50698 50704 12:40293894_C_CAT 50606-50618, 50640-50647, 50661, 50663- rs1491426750_REF 50620-50628, 50649-50651, 50670, 50672- 50630-50639 50653-50660 50684 12:40293894_C_CAT 50607, 50616, 50640, 50645, 50661, 50666, rs1491426750_SNP 50618, 50620- 50647, 50649, 50668, 50670, 50621, 50688, 50658, 50695- 50682, 50700- 50690, 50692- 50698, 50729- 50702, 50704, 50693, 50705- 50736 50737-50746 50728 12:40293896_C_T 50606-50612, 50640-50642, 50661, 50663- rs5006481_REF 50614-50618, 50644-50647, 50670, 50672- 50620-50622, 50649-50651, 50674, 50677- 50624-50628, 50654-50658, 50678, 50680- 50630-50631, 50660 50682, 50684 50634-50639 12:40293896_C_T 50607, 50616, 50640, 50645, 50661, 50666, rs5006481_SNP 50618, 50620- 50647, 50649, 50668, 50670, 50621, 50685- 50658, 50696- 50682, 50699, 50687, 50689- 50698 50701-50704 50694 12:40294132_CATCTATCTAT 50747-50753 50754-50755 CT_C rs139565575_REF 12:40294132_CATCTATCT_C 50747-50753 50754-50755 rs150276517_REF 12:40294132_CATCT_C 50747-50753 50754-50755 rs72446556_REF 12:40294132_C_CATCT 50747-50753 50754-50755 rs1186032972_REF 12:40294279_A_G 50756-50802 50803-50852 50853-50903 rs12820920_REF 12:40294279_A_G 50756-50757, 50803-50804, 50854, 50857, rs12820920_SNP 50759-50760, 50807, 50813, 50863, 50865- 50767-50768, 50815-50816, 50866, 50869, 50773, 50776, 50821, 50829, 50880, 50888, 50781, 50793, 50838, 50843, 50893, 50898, 50904-50941 50942-50981 50982-51023 12:40294893_T_C 51024-51034 51035-51040 51041-51048 rs7966550_REF 12:40294893_T_C 51024-51025, 51035-51036, 51041, 51043- rs7966550_SNP 51028-51034, 51038-51040, 51044, 51046, 51049-51056 51057-51061 51048, 51062- 51069 12:40295119_A_G 51070-51087 51088-51089 rs12314692_SNP 12:40295786_C_T 51090-51136 51137-51184 51185-51236 rs11175922_REF 12:40295786_C_T 51091, 51096, 51138, 51141, 51185, 51187, rs11175922_SNP 51098, 51100, 51144, 51146, 51190, 51194, 51103-51105, 51148, 51151- 51196, 51201, 51118, 51125- 51152, 51166, 51216, 51222, 51126, 51237- 51173-51174, 51224-51225, 51268 51269-51296 51297-51330 12:40296087_T_C 51331-51357 51358-51376 51377-51403 rs73102746_REF 12:40296087_T_C 51335, 51340, 51360, 51362, 51377, 51380, rs73102746_SNP 51346-51347, 51366, 51370, 51385, 51390, 51353, 51404- 51438-51466 51395, 51467- 51437 51499 12:40296180_T_C 51500-51536 51537-51568 51569-51604 rs10784497_REF 12:40296180_T_C 51514, 51524, 51559, 51563, 51598, 51600, rs10784497_SNP 51530, 51533, 51565, 51642- 51602, 51677- 51605-51641 51676 51713 12:40296259_A_G 51714-51749 51750-51773 51774-51799 rs17443815_REF 12:40296259_A_G 51716, 51726- 51751, 51759- 51775, 51778, rs17443815_SNP 51727, 51737, 51760, 51768, 51785, 51793, 51747, 51800- 51771, 51838- 51797, 51873- 51837 51872 51912 12:40296904_C_A 51913-51959 51960-52005 52006-52057 rs7957057_REF 12:40296904_C_A 51914-51915, 51961-51962, 52007-52008, rs7957057_SNP 51923-51924, 51967, 51970- 52014, 52017- 51931, 51934, 51971, 51977, 52018, 52025, 51937, 51941, 51987, 51993- 52035, 52038, 51947-51948, 51994, 52001, 52050, 52056, 52058-52087 52088-52115 52116-52147 12:40297067_CTT_C 52148-52193 52194-52243 52244-52295 rs76607433_REF 12:40297067_CTT_C 52163-52165, 52212-52213, 52256, 52264, rs76607433_SNP 52167, 52172, 52215, 52220, 52266, 52279- 52178, 52187, 52227-52228, 52280, 52291, 52189, 52296- 52239, 52242, 52294-52295, 52332 52333-52371 52372-52412 12:40297142_G_A 52413-52452 52453-52478 52479-52511 rs28365224_REF 12:40297142_G_A 52420, 52423, 52459, 52466, 52487, 52490, rs28365224_SNP 52427, 52435, 52472, 52474, 52494, 52503, 52441, 52450, 52477, 52523- 52506, 52510, 52512-52522 52525 52526-52534 12:40298741_C_T 52535-52541 52542-52553 52554-52566 rs7962011_SNP 12:40299423_A_C 52567-52572 52573-52574 52575-52577 rs10878336_REF 12:40299423_A_C 52567-52572, 52573-52574, 52575-52577, rs10878336_SNP 52578-52588 52589-52590 52591-52595 12:40299999_C_G 52596-52643 52644-52693 52694-52745 rs11175934_REF 12:40299999_C_G 52605, 52610, 52653, 52658, 52703, 52708- rs11175934_SNP 52620, 52630, 52668, 52678- 52709, 52729- 52632, 52635, 52679, 52684, 52730, 52735, 52640-52643, 52686, 52690- 52737, 52742- 52746-52783 52691, 52693, 52743, 52745, 52784-52823 52824-52865 12:40300004_G_T 52596-52599, 52644-52647, 52694-52697, rs11175935_REF 52601-52604, 52649-52652, 52699-52702, 52606-52607, 52654-52655, 52704-52705, 52609-52610, 52657-52658, 52707, 52709, 52614-52619, 52662-52667, 52713-52728, 52621-52634, 52669-52677, 52730-52734, 52636-52640, 52679-52683, 52736-52741, 52643, 52866- 52685-52690, 52743-52745, 52874 52692-52693, 52881-52887 52875-52880 12:40300004_G_T 52615, 52618, 52663, 52671, 52719, 52722, rs11175935_SNP 52623, 52625, 52673, 52687, 52724, 52738, 52637, 52869- 52692, 52878, 52744, 52884, 52870, 52873- 52926-52964 52887, 52965- 52874, 52888- 53005 52925 12:40300216_T_C 53006-53042 53043-53076 53077-53112 rs17491061_REF 12:40300216_T_C 53009, 53013, 53046, 53049, 53080, 53093, rs17491061_SNP 53021, 53028- 53058, 53063- 53098-53099, 53029, 53040, 53064, 53147- 53105, 53181- 53113-53146 53180 53218 12:40300597_G_A 53219-53266 53267-53316 53317-53368 rs11175941_REF 12:40300597_G_A 53224, 53226, 53270, 53273, 53320, 53323, rs11175941_SNP 53238-53239, 53275, 53287- 53327, 53338- 53245-53246, 53288, 53294- 53339, 53345- 53250-53251, 53295, 53299, 53346, 53360, 53255, 53260, 53309, 53312, 53363, 53366, 53369-53406 53407-53446 53447-53488 12:40301293_G_A 53489 53490-53492 53493-53499 rs10878340_SNP 12:40303632_G_A 53500-53533 53534-53557 53558-53589 rs10784498_REF 12:40303632_G_A 53500, 53512, 53547, 53607- 53565, 53573, rs10784498_SNP 53525-53526, 53612 53582, 53613- 53590-53606 53622 12:40303675_C_T 53623-53670 53671-53715 53716-53763 rs10878341_REF 12:40303675_C_T 53626, 53633, 53674, 53681, 53719, 53731, 53686, 53703, rs10878341_SNP 53638, 53648, 53707, 53802- 53750, 53753, 53650, 53655, 53841 53756, 53762, 53659, 53662, 53842-53883 53668-53669, 53764-53801 12:40304688_C_T 53884-53915 53916-53949 53950-53988 rs10784499_REF 12:40304688_C_T 53886-53887, 53917, 53919, 53951, 53953, rs10784499_SNP 53892, 53896, 53925, 53927, 53959, 53962, 53903, 53909, 53929, 53937, 53964, 53978, 53913, 53989- 53943, 53947, 53981-53982, 54023 54024-54043 53986, 54044- 54072 12:40304884_A_G 54073 54074 rs17484286_REF 12:40304884_A_G 54073 54074 rs17484286_SNP 12:40306303_CTT_C 54075-54120 54121-54144 54145-54177 rs146071397_REF 12:40306303_CTT_C 54085, 54093, 54131, 54136, 54166, 54175, rs146071397_SNP 54101-54102, 54144, 54216- 54177, 54250- 54107, 54116, 54249 54288 54178-54215 12:40306782_C_T 54289-54325 54326-54357 54358-54391 rs10878343_REF 12:40306782_C_T 54392-54421 54422-54443 54444-54471 rs10878343_SNP 12:40307469_G_T 54472-54488 54489-54493 54494-54501 rs11564180_REF 12:40307469_G_T 54473, 54481, 54501 rs11564180_SNP 54502-54504 12:40307586_G_A 54505-54510 rs11175958_REF 12:40307927_A_G 54511-54535 54536-54562 54563-54591 rs11175959_REF 12:40307927_A_G 54511, 54513, 54536-54537, 54563-54565, rs11175959_SNP 54518, 54532, 54539, 54544, 54567, 54572, 54534, 54592- 54558, 54612- 54634-54657 54611 54633 12:40308029_C_T , 54658-54659 54660-54663 rs113772607_SNP 12:40308362_C_T 54664-54697 54698-54726 54727-54758 rs11564149_REF 12:40308362_C_T 54670, 54686, 54702, 54717, 54732, 54747, rs11564149_SNP 54688, 54690, 54784-54805 54749, 54806- 54697, 54759- 54829 54783 12:40308969_C_T 54830-54876 54877-54924 54925-54976 rs11175963_REF 12:40308969_C_T 54830, 54832, 54890, 54893, 54934, 54940, rs11175963_SNP 54844, 54848, 54895, 54906- 54943, 54945, 54859, 54861, 54907, 54909, 54956-54957, 54863-54864, 54912-54913, 54959, 54962, 54867, 54869, 55009-55024 54968, 54970, 54977-55008 55025-55051 12:40309044_A_T 55052-55063 55064-55073 55074-55085 rs7305344_REF 12:40309044_A_T 55052-55053, 55064-55065, 55074-55075, rs7305344_SNP 55055, 55058- 55067-55068, 55077-55078, 55059, 55086- 55070, 55093- 55083, 55098- 55092 55097 55104 12:40309109_G_A 55105-55140 55141-55181 55182-55229 rs7133914_REF 12:40309109_G_A 55120, 55123, 55149, 55162- 55191, 55204- rs7133914_SNP 55126, 55230- 55164, 55176, 55207, 55211, 55260 55261-55287 55288-55318 12:40309185_G_A 15754, 15758, 15800, 15806, 15849, 15851, rs11175964_REF 15769, 15786, 15817, 15835, 15869, 15887, 15903, 15910, 15845, 15949, 15897, 15999, 15916, 55319- 15956, 15963, 16003, 16007, 55354 15975, 55355- 16014, 16026, 55395 55396-55437 12:40309185_G_A 55332, 55336, 55357, 55362, 55397, 55401, rs11175964_SNP 55339, 55341- 55366, 55371, 55406, 55411, 55342, 55350- 55376, 55381, 55416, 55424, 55351, 55354, 55387, 55391- 55428, 55433, 55438-55475 55392, 55395, 55436-55437, 55476-55515 55516-55557 12:40309285_C_CGT 55558-55577 55578-55596 55597-55617 rs397805328_REF 12:40309350_C_A 55618-55632 55633-55637 55638-55645 rs11175966_REF 12:40309350_C_A 55618, 55622- 55633, 55635 55638, 55640, rs11175966_SNP 55623, 55631- 55642 55632, 55646 12:40310063_A_G 55647-55652 55653-55657 55658-55663 rs10784501_REF 12:40310063_A_G 55647-55649, 55653-55657, 55658-55660, rs10784501_SNP 55651-55652, 55675-55677 55662-55663, 55664-55674 55678-55682 12:40310755_T_C 55683-55725 55726-55766 55767-55811 rs41286474_REF 12:40310755_T_C 55688-55689, 55730, 55732- 55771, 55773, rs41286474_SNP 55693-55694, 55733, 55737, 55778, 55781, 55697-55698, 55740, 55745, 55785, 55790, 55702, 55705, 55748, 55758, 55801-55803, 55717, 55812- 55850-55887 55888-55929 55849 12:40311346_CT_C 55930-55947 55948-55958 55959-55971 rs869212703_REF 12:40311346_CT_C 55940, 55945 55969 rs869212703_SNP 12:40311356_T_A 55940, 55945, 55977 55969, 55978- rs2069228_REF 55972-55976 55979 12:40311356_T_A 55940, 55945, 55977 55969, 55978- rs2069228_SNP 55972-55976 55979 12:40311898_C_G 55980-55989 55990-55993 55994-56000 rs10784503_REF 12:40311898_C_G 55981, 55983, 55990-55991, 55995-55996, rs10784503_SNP 55985, 55987, 55993, 56006 55998-55999, 55989, 56001- 56005 56007-56009 12:40311964_G_GT 56010-56028 56029-56044 56045-56062 rs386376250_REF 12:40311964_G_GT 56027 56040 56061 rs386376250_SNP 12:40312033_C_T 56063-56100 56101-56134 56135-56172 rs11564128_REF 12:40312033_C_T 56073, 56080, 56108, 56111, 56140-56141, rs11564128_SNP 56082, 56090- 56114, 56116, 56145, 56149, 56091, 56094, 56123, 56130, 56161, 56168, 56173-56194 56195-56202 56203-56218 12:40312573_G_T 56219-56265 56266-56312 56313-56362 rs2896975_REF 12:40312573_G_T 56219, 56229, 56270, 56277, 56314, 56318, rs2896975_SNP 56243-56244, 56292, 56299, 56325, 56340, 56251, 56257, 56305, 56309, 56342, 56358, 56260, 56263- 56312, 56401- 56361-56362, 56265, 56363- 56400 56440 56441-56482 12:40312729_G_A 56483-56530 56531-56580 56581-56632 rs7302503_REF 12:40312729_G_A 56484, 56488, 56532, 56541, 56591, 56596, rs7302503_SNP 56493, 56498, 56546, 56548, 56598, 56600, 56501, 56519, 56568, 56570, 56619-56620, 56521, 56528- 56577-56580, 56629-56632, 56530, 56633- 56671-56710 56711-56752 56670 12:40312802_T_C 56753-56766 56767-56775 56776-56787 rs7137160_REF 12:40312802_T_C 56753, 56755, 56767-56769, 56776-56777, rs7137160_SNP 56757-56758, 56771-56775, 56779, 56781- 56760-56761, 56806-56813 56782, 56784- 56763-56766, 56787, 56814- 56788-56805 56827 12:40312938_TTAAGAGA_T 56828-56858 56859-56900 56901-56948 rs369794148_REF 12:40312938_TTAAGAGA_T 56829, 56833- 56860, 56867, 56901, 56904, rs369794148_SNP 56834, 56848, 56884, 56896, 56911, 56921, 56857, 56949- 56987-57026 56944, 57027- 56986 57068 12:40312990_GA_G 57069-57109 57110-57150 57151-57196 rs34709132_REF 12:40312990_GA_G 57070-57071, 57110, 57112- 57151, 57154- rs34709132_SNP 57075-57077, 57113, 57117, 57155, 57160, 57088, 57091, 57120, 57133, 57163, 57165, 57094, 57100, 57136, 57141, 57184-57186, 57197-57228 57229-57260 57261-57296 12:40313399_A_G 57297-57338 57339-57369 57370-57409 rs10467144_REF 12:40313399_A_G 57309, 57314, 57345, 57350, 57378, 57380, rs10467144_SNP 57317, 57321, 57353, 57362, 57385, 57397, 57327, 57334, 57364, 57367, 57402, 57405, 57410-57447 57448-57486 57409, 57487- 57527 12:40313476_C_T 57528-57572 57573-57618 57619-57668 rs11175985_REF 12:40313476_C_T 57534, 57545, 57578, 57591, 57624-57625, rs11175985_SNP 57547-57548, 57593, 57596- 57640, 57642, 57551, 57565, 57597, 57611, 57646-57647, 57570, 57669- 57616, 57706- 57661, 57666, 57705 57738 57739-57775 12:40313594_T_G 57776-57801 57802-57815 57816-57836 rs10784504_REF 12:40313594_T_G 57779, 57781, 57804, 57808, 57819-57820, rs10784504_SNP 57784-57785, 57811-57812, 57826, 57829- 57792-57793, 57869-57891 57830, 57892- 57795, 57798- 57919 57799, 57837- 57868 12:40313687_G_GT 57920-57926 57927-57928 57929-57930 rs57421387_REF 12:40313687_G_GT 57923 57927 57930 rs57421387_SNP 12:40315043_A_T 57931-57946 57947-57954 57955-57964 rs7973058_REF 12:40315043_A_T 57933, 57935- 57947-57948, 57956-57957, rs7973058_SNP 57936, 57939, 57951, 57953- 57960-57961, 57945, 57965- 57954, 57976- 57964, 57979- 57975 57978 57983 12:40315090_T_C 57984-58030 58031-58080 58081-58132 rs1427267_REF 12:40315090_T_C 57984-57985, 58032, 58034- 58082, 58084- rs1427267_SNP 57987-57988, 58035, 58044, 58085, 58093, 57992, 57996, 58047, 58056- 58095, 58108, 57999, 58008- 58057, 58059, 58110, 58117, 58009, 58011, 58074, 58076, 58126, 58128, 58133-58168 58169-58208 58209-58250 12:40315120_C_T 58251-58296 58297-58344 58345-58396 rs1427266_REF 12:40315120_C_T 58253, 58257- 58297, 58299, 58345-58346, rs1427266_SNP 58258, 58267- 58303, 58305- 58349, 58353, 58268, 58276, 58306, 58314- 58355, 58366, 58279-58280, 58315, 58323, 58374, 58377, 58284, 58289, 58327, 58332, 58379, 58384, 58397-58425 58426-58453 58454-58487 12:40315383_A_G 58488-58518 58519-58544 58545-58582 rs721713_REF 12:40315383_A_G 58494, 58498, 58524, 58526- 58547, 58553, rs721713_SNP 58500, 58502, 58527, 58529, 58560, 58562, 58504, 58510, 58531, 58536, 58564, 58572, 58512-58513, 58538-58539, 58574-58575, 58516, 58518, 58542, 58544, 58580, 58582, 58583-58620 58621-58652 58653-58688 12:40315516_G_A 58689-58735 58736-58785 58786-58837 rs721712_REF 12:40315516_G_A 58694, 58697- 58741, 58744, 58791, 58793, rs721712_SNP 58698, 58704, 58751, 58756, 58795, 58802, 58709, 58712, 58759-58760, 58810, 58816, 58717, 58723, 58765, 58771, 58818, 58823, 58727, 58732, 58775, 58781, 58827, 58833, 58838-58875 58876-58915 58916-58957 12:40315526_G_A 58689-58693, 58736-58740, 58786-58790, rs721711_REF 58695-58699, 58742-58746, 58792-58798, 58702-58703, 58749-58750, 58800-58801, 58705, 58710- 58752, 58757- 58803, 58808- 58712, 58715, 58760, 58763, 58811, 58814, 58717, 58719- 58765, 58767- 58816, 58819- 58720, 58722, 58768, 58770, 58820, 58822, 58725, 58727- 58773-58775, 58825-58827, 58728, 58731, 58777, 58780, 58829, 58832, 58733-58735, 58783-58785, 58835-58837, 58958-58977 58978-58997 58998-59017 12:40315526_G_A 58695, 58710- 58742, 58757- 58792, 58798, rs721711_SNP 58711, 58728, 58758, 58774, 58808-58809, 58733, 58963, 58783, 58983, 58826, 59001, 58968, 58974- 58988, 58991, 59003, 59008, 58975, 58977, 58995, 58997, 59011, 59017, 59018-59051 59052-59081 59082-59119 12:40315611_T_A 59120-59160 59161-59200 59201-59245 rs721709_REF 12:40315611_T_A 59126, 59139, 59167, 59182- 59208, 59225, rs721709_SNP 59142-59144, 59183, 59190, 59232, 59234, 59151, 59246- 59196, 59281- 59240, 59244, 59280 59315 59316-59354 12:40315697_T_G 59355-59399 59400-59443 59444-59490 rs4768227_REF 12:40315697_T_G 59361, 59374- 59403, 59407, 59445, 59448, 59375, 59377, 59420, 59424, 59452, 59465- 59379, 59395, rs4768227_SNP 59399, 59491- 59440, 59529- 59466, 59470, 59528 59568 59569-59610 12:40315806_AT_A 59611-59612 rs398098159_REF 12:40315806_AT_A 59611-59617 59618-59619 rs398098159_SNP 12:40315822_G_T 59611-59613, 59642-59650 59651-59663 rs4767969_REF 59620-59641 12:40315822_G_T 59620-59621, 59667-59668 rs4767969_SNP 59631, 59638, 59664-59666 12:40316155_G_A 59669-59710 59711-59753 59754-59798 rs10878356_REF 12:40316155_G_A 59670, 59675, 59712, 59718, 59755, 59762- rs10878356_SNP 59694-59695, 59737-59738, 59763, 59782, 59799-59835 59836-59870 59871-59907 12:40317268_C_T 59908-59917 59918-59922 59923-59932 rs10784506_REF 12:40317268_C_T 59911-59912, 59918, 59921- 59924, 59928- rs10784506_SNP 59915 59922 59930, 59932 12:40317289_G_A 59908, 59911- 59918-59919, 59924, 59926, rs10878358_REF 59913, 59915- 59921-59922, 59928-59932, 59916, 59933- 59961-59970 59971-59987 59960 12:40317289_G_A 59908, 59913, 59919, 59998- 59926, 59931, rs10878358_SNP 59916, 59950, 60001 60002-60007 59955, 59988- 59997 12:40317292_A_T 59933-59960, 59918-59919, 59924, 59926, rs7132073_REF 60008-60009 59961-59970, 59928, 59931, 60010 59971-59987, 60011 12:40317292_A_T 59950, 59958, 59919, 59970, 59926, 59931, rs7132073_SNP 59960, 60012- 60039-60049 59975, 59977, 60038 59979, 60050- 60066 12:40318196_C_A 60067-60112 60113-60157 60158-60204 rs1427265_REF 12:40318196_C_A 60068, 60071, 60116, 60126, 60162, 60172, rs1427265_SNP 60075, 60078, 60129, 60131, 60175, 60191, 60083, 60085, 60147, 60243- 60194, 60282- 60095, 60101, 60281 60322 60107, 60205- 60242 12:40318557_C_T 60323-60368 60369-60407 60408-60455 rs17466339_REF 12:40318557_C_T 60328, 60330, 60372, 60384- 60415-60416, rs17466339_SNP 60338, 60341, 60385, 60391, 60429-60430, 60348, 60355, 60397, 60401, 60436, 60443, 60359, 60456- 60481-60499 60447, 60500- 60480 60522 12:40318773_A_C 60523-60548 60549-60555 60556-60568 rs2114569_REF 12:40318773_A_C 60524, 60526- 60549, 60552, 60556, 60559, rs2114569_SNP 60527, 60529- 60605-60628 60566, 60629- 60530, 60533- 60662 60535, 60541- 60542, 60569- 60604 12:40318818_G_A 60663-60710 60711-60759 60760-60811 rs2114568_REF 12:40318818_G_A 60667, 60670, 60735, 60742- 60783, 60785, rs2114568_SNP 60687, 60694- 60743, 60746, 60792-60793, 60695, 60698- 60751, 60753- 60802, 60804- 60699, 60705- 60755, 60758, 60806, 60809- 60706, 60709, 60850-60888 60810, 60889- 60812-60849 60929 12:40319201_G_A 60930-60967 60968-61002 61003-61043 rs2162469_REF 12:40319201_G_A 60943, 61044- 61075-61084 61085-61100 rs2162469_SNP 61074 12:40319398_G_A 61101-61148 61149-61198 61199-61250 rs10784509_REF 12:40319398_G_A 61107, 61118- 61155, 61166- 61216, 61224- rs10784509_SNP 61119, 61122, 61167, 61174- 61229, 61232, 61126-61127, 61177, 61180, 61236, 61246, 61130, 61134, 61184, 61194, 61329-61370 61144, 61146, 61289-61328 61251-61288 12:40319758_T_G 61371-61417 61418-61457 61458-61504 rs1896254_REF 12:40319758_T_G 61371, 61375, 61418, 61421, 61458, 61461, rs1896254_SNP 61378, 61383, 61427, 61431, 61487-61488, 61387, 61405, 61445, 61449, 61493, 61498, 61410, 61413, 61543-61582 61501, 61583- 61416, 61505- 61624 61542 12:40319784_CA_C 61625-61627 61628-61630 61631-61634 rs5797667_REF 12:40319784_CA_C 61626 61629 61632, 61634 rs5797667_SNP 12:40319786_AT_A 61626 61629 61632, 61634 rs373676201_REF 12:40319793_T_C 61635-61636 61637-61638 61639-61640 rs1896253_REF 12:40319793_T_C 61635-61636 61637-61638 61639-61640 rs1896253_SNP 12:40319957_T_C 61641-61644 61645 rs1896252_REF 12:40319957_T_C 61641-61644, 61647 61645, 61648- rs1896252_SNP 61646 61650 12:40320032_C_A 19471, 19475, 19516-19517, 19558-19559, rs1427263_REF 19477, 19486, 19520, 19522, 19561, 19564, 19491, 19620, 19530, 19644, 19574, 19662, 19622, 61651- 19646-19647, 19672, 19674- 61688 61689-61730 19675, 61731- 61773 12:40320032_C_A 61657, 61660, 61692, 61698, 61733, 61739, rs1427263_SNP 61665, 61668, 61703, 61706, 61747, 61756, 61676, 61678, 61716, 61718, 61758, 61760, 61683, 61686- 61723, 61728- 61765, 61770, 61688, 61774- 61730, 61812- 61772-61773, 61811 61851 61852-61893 12:40320071_A_G 61894-61898 61899-61900 61901-61904 rs11176013_REF 12:40320071_A_G 61894-61898, 61899-61900 61901-61904, rs11176013_SNP 61905-61911 61912 12:40320099_T_A 19709, 19712, 19739, 19752, 19779, 19790, rs11564148_REF 19823, 19825, 19867, 19880, 19793-19794, 19838, 19841, 19884, 19888, 19913, 19920, 19958, 19966, 20001, 20009, 19930, 19934, 19969, 19974, 20017, 20024, 20037, 20047, 19979, 20087, 20029, 20129, 20070, 20075, 20093, 20106, 20134, 20139, 20077, 20173, 20116, 61913- 20152, 20161, 20178, 20184, 61931 61932-61951 20199, 20207, 61952-61977 12:40320099_T_A 19958, 19966, 20001, 20009, 20037, 20047, rs11564148_SNP 19969, 19974, 20017, 20024, 20070, 20075, 19979, 61917, 20029, 62014- 20077, 61966, 61929, 61931, 62044 61969, 62045- 61978-62013 62081 12:40320207_A_G 62082-62103 62104-62115 62116-62131 rs11564205_REF 12:40320207_A_G 62084, 62092, 62165-62187 62188-62212 rs11564205_SNP 62132-62164 12:40320386_G_A 62213-62228 62229-62241 62242-62257 rs10748030_REF 12:40320386_G_A 62215, 62221, 62231, 62234, 62244, 62247, rs10748030_SNP 62223, 62225- 62236, 62239, 62249, 62251, 62226, 62258- 62265-62268 62255, 62269- 62264 62274 12:40320464_T_C 62275-62284 62285-62296 62297-62310 rs10748031_REF 12:40320464_T_C 62275-62277, 62285-62286, 62298-62300, rs10748031_SNP 62280-62281, 62288, 62290, 62303, 62305, 62284, 62311- 62292-62293, 62307, 62310, 62336 62295, 62337- 62362-62386 62361 12:40320465_G_T 62275-62283, 62285-62291, 62297-62299, rs10748032_REF 62387-62388 62293-62296, 62301-62310, 62389-62390 62391-62392 12:40320465_G_T 62275-62277, 62286, 62288, 62298-62299, rs10748032_SNP 62280-62281, 62290, 62293, 62305, 62307, 62387, 62393- 62295, 62390, 62310, 62392, 62419 62420-62446 62447-62473 12:40320530_T_C 62474-62518 62519-62558 62559-62601 rs11564177_REF 12:40320530_T_C 62476, 62481, 62531, 62545, 62571, 62587, rs11564177_SNP 62489, 62506, 62548, 62640- 62590, 62598, 62513-62514, 62679 62680-62721 62518, 62602- 62639 12:40320593_C_T 62722-62762 62763-62790 62791-62828 rs10748033_REF 12:40320593_C_T 62724, 62726, 62764, 62768, 62792, 62797, rs10748033_SNP 62735-62736, 62772-62773, 62802-62803, 62746, 62753, 62785, 62848- 62805, 62822, 62829-62847 62860 62861-62877 12:40320699_T_G 62878-62895 62896-62913 62914-62933 rs10735931_REF 12:40320699_T_G 62880-62881, 62899-62900, 62915, 62918, rs10735931_SNP 62884, 62888, 62902, 62906, 62920, 62926, 62892, 62934- 62910, 62953- 62930, 62968- 62952 62967 62984 12:40320816_C_T 62985-63024 63025-63058 63059-63102 rs10748034_REF 12:40320816_C_T 62994, 63003, 63033, 63044, 63068, 63072, rs10748034_SNP 63008, 63012- 63047, 63053, 63090, 63096, 63013, 63019, 63055, 63126- 63098, 63100, 63103-63125 63142 63143-63163 12:40320983_A_T 63164-63209 63210-63255 63256-63306 rs10878368_REF 12:40320983_A_T 63176, 63179- 63217, 63223, 63263, 63269, rs10878368_SNP 63181, 63190- 63226-63228, 63272, 63274, 63191, 63199- 63237-63238, 63283-63284, 63200, 63202- 63246, 63249, 63293, 63298- 63203, 63307- 63344-63380 63300, 63381- 63343 63421 12:40321211_T_A 63422-63424 63425-63427 63428-63431 rs7307276_REF 12:40321211_T_A 63422-63424 63425-63427 63428-63431 rs7307276_SNP 12:40321446_T_C 63432-63479 63480-63529 63530-63581 rs4768229_REF 12:40321446_T_C 63436, 63440- 63484, 63488, 63532, 63535, rs4768229_SNP 63441, 63450, 63493, 63499, 63539, 63544, 63461, 63464, 63506, 63511, 63553, 63558, 63467-63468, 63517-63518, 63569-63570, 63477-63478, 63527-63528, 63579-63580, 63582-63619 63620-63659 63660-63701 12:40321500_G_A 63702-63731 63732-63761 63762-63793 rs4768230_REF 12:40321500_G_A 63711, 63720, 63739, 63741, 63769, 63771, rs4768230_SNP 63725, 63728, 63750, 63758, 63778, 63781, 63730, 63794- 63760, 63817- 63791, 63838- 63816 63837 63860 12:40321508_C_CA 63704-63705, 63734-63735, 63764-63765, rs397968969_REF 63707-63709, 63737-63738, 63767-63768, 63712-63713, 63742-63743, 63770, 63772- 63716, 63721- 63746, 63751- 63773, 63776, 63722, 63726- 63752, 63754, 63782-63783, 63727, 63729, 63756-63757, 63785, 63787- 63731, 63861- 63759, 63761, 63788, 63790, 63863 63864-63867 63792-63793, 63868-63871 12:40321508_C_CA 63709, 63713, 63735, 63743, 63765, 63768, rs397968969_SNP 63862-63863 63864-63866 63773, 63868- 63869, 63871 12:40321575_G_A 63872-63909 63910-63943 63944-63980 rs11564204_REF 12:40321575_G_A 63873, 63875, 63911, 63917, 63945, 63947, rs11564204_SNP 63882, 63892, 63933, 64010- 63953, 63956, 63899, 63981- 64038 64039-64069 64009 12:40321788_C_T 64070-64114 64115-64157 64158-64203 rs4423250_REF 12:40321788_C_T 64083, 64086, 64128, 64134, 64171, 64177, rs4423250_SNP 64089, 64091, 64136, 64241- 64192, 64194, 64102, 64105- 64266 64267-64301 64106, 64204- 64240 12:40322213_C_T 64302-64321 64322-64323 64324-64329 rs7137665_REF 12:40322458_T_C 21565, 21569, 21580, 64351- 21581, 21583, rs10878371_REF 21573, 21575, 64365 64366-64385 21577, 64330- 64350 12:40322458_T_C 64386-64422 64423-64454 64455-64488 rs10878371_SNP 12:40322672_T_C 64489-64490 rs11176022_REF 12:40322672_T_C 64489-64494 64495-64496 rs11176022_SNP 12:40322834_CAT_C 64497-64542 64543-64589 64590-64641 rs60750674_REF 12:40322834_CAT_C 64498, 64500, 64543, 64545, 64591, 64593, rs60750674_SNP 64506-64507, 64551-64552, 64601, 64609, 64516-64517, 64560, 64562, 64611, 64622, 64526, 64531, 64572, 64578, 64628, 64633, 64642-64680 64681-64721 64722-64764 12:40322892_A_G 64765-64812 64813-64860 64861-64912 rs10878372_REF 12:40322892_A_G 64770, 64772, 64818, 64820, 64866, 64868, rs10878372_SNP 64775, 64784, 64823, 64831, 64880-64881, 64786, 64789, 64833, 64836, 64883, 64886- 64793-64794, 64840, 64852, 64887, 64891, 64803, 64811, 64856, 64859, 64903, 64908, 64913-64950 64951-64990 64991-65032 12:40323798_CTTT_C 65033-65044 65045-65056 65057-65070 rs398055492_REF 12:40323798_CTT_C 65033-65044 65045-65056 65057-65070 rs761931834_REF 12:40323798_CT_C 65033-65044 65045-65056 65057-65070 rs1415940047_REF 12:40324198_G_A 65071-65106 65107-65140 65141-65176 rs17466430_REF 12:40324198_G_A 65080, 65086, 65116, 65121, 65150, 65152, rs17466430_SNP 65090, 65101, 65124, 65126, 65159, 65161, 65104, 65177- 65136, 65204- 65171, 65231- 65203 65230 65259 12:40324651_A_T 65260-65288 65289-65300 65301-65316 rs11176030_REF 12:40324651_A_T 65264, 65268, 65342-65353 65354-65369 rs11176030_SNP 65273, 65279, 65286, 65317- 65341 12:40325081_T_C 65370-65377 65378-65387 65388-65399 rs3747891_REF 12:40325081_T_C 65371, 65373, 65379-65380, 65389, 65397, rs3747891_SNP 65400-65421 65422-65447 65448-65477 12:40325698_A_C 65478-65509 65510-65537 65538-65569 rs7298930_REF 12:40325698_A_C 65480, 65483, 65512, 65514, 65542, 65544- rs7298930_SNP 65488, 65491, 65516-65517, 65545, 65552, 65493, 65496- 65523, 65527- 65556-65558, 65497, 65499- 65528, 65530- 65561-65562, 65501, 65570- 65532, 65602- 65566, 65630- 65601 65629 65665 12:40326004_G_C 65666-65713 65714-65763 65764-65815 rs17444089_REF 12:40326004_G_C 65669, 65672, 65717, 65720, 65767, 65772, rs17444089_SNP 65674, 65680, 65722, 65728, 65778, 65780, 65682, 65687, 65730, 65737, 65787, 65800, 65689, 65692, 65742, 65750, 65802, 65808, 65694, 65702, 65752, 65758, 65810, 65812, 65816-65853 65854-65893 65894-65935 12:40326202_T_C 65936 65937-65939 65940-65944 rs10878377_REF 12:40326202_T_C 65936 65937-65938, 65940-65942, rs10878377_SNP 65945 65946-65947 12:40326497_A_C 65948-65959 65960-65971 65972-65983 rs12316801_REF 12:40326497_A_C 65952-65954, 65965-65966, 65976, 65978, rs12316801_SNP 65958-65959, 65968, 65970- 65980, 65982- 65984-65994 65971, 65995- 65983, 66006- 66005 66018 12:40326499_A_C 65948-65959, 65960-65971, 65972-65983, rs17444096_REF 66019-66022 66023-66026 66027-66030 12:40326499_A_C 65954, 65958- 65968, 65970- 65976, 65980, rs17444096_SNP 65959, 66021- 65971, 66025- 65983, 66029- 66022, 66031- 66026, 66044- 66030, 66057- 66043 66056 66069 12:40327240_G_A 66070-66116 66117-66166 66167-66218 rs11564175_REF 12:40327240_G_A 66077, 66082, 66125, 66130, 66175, 66180, rs11564175_SNP 66084, 66099, 66132, 66147, 66182, 66189, 66101, 66104- 66152-66154, 66198, 66203, 66105, 66109, 66158, 66164, 66205-66206, 66112, 66116, 66166, 66257- 66210, 66216, 66219-66256 66296 66297-66338 12:40328650_A_G 66339-66385 66386-66431 66432-66482 rs2404832_REF 12:40328650_A_G 66344, 66350, 66398, 66400, 66444, 66447, rs2404832_SNP 66353, 66369, 66414, 66417, 66458, 66462, 66371-66372, 66419-66420, 66467-66468, 66380-66381, 66428, 66431, 66474, 66477- 66383, 66385, 66521-66560 66478, 66561- 66483-66520 66602 12:40328747_C_A 66603-66649 66650-66695 66696-66747 rs17466486_REF 12:40328747_C_A 66616, 66618- 66664-66665, 66701, 66713, rs17466486_SNP 66619, 66621, 66667, 66677, 66715-66716, 66624, 66631, 66680-66681, 66718, 66727, 66634-66635, 66685, 66690, 66730-66731, 66639, 66748- 66783-66807 66736, 66741, 66782 66808-66836 12:40328922_C_T 66837-66883 66884-66931 66932-66981 rs11564267_REF 12:40328922_C_T 66841, 66844- 66888, 66892, 66936, 66939, rs11564267_SNP 66845, 66850, 66897, 66900, 66949, 66961, 66853, 66861, 66914, 66922, 66964, 66972, 66867, 66871, 66928, 66931, 66978, 66981, 66875, 66881, 67020-67059 67060-67101 66982-67019 12:40329067_AG_A 67102-67126 67127-67132 67133-67146 rs17520244_REF 12:40329067_AG_A 67108-67109, 67130 67135, 67139, rs17520244_SNP 67113, 67121, 67144 67147-67148 12:40329673_T_A 67149-67158 67159-67160 67161-67163 rs10506152_REF 12:40329673_T_A 67150, 67152, 67159-67160 67161-67163 rs10506152_SNP 67155, 67157, 67164-67169 12:40330098_C_T 67170-67212 67213-67255 67256-67300 rs11176052_REF 12:40330098_C_T 67173, 67183, 67226, 67245, 67266, 67270, rs11176052_SNP 67199, 67202, 67254, 67339- 67290, 67378- 67207, 67211, 67377 67418 67301-67338 12:40330228_C_T 67419-67460 67461-67489 67490-67525 rs11176053_REF 12:40330228_C_T 67427, 67436, 67463, 67469, 67490, 67494, rs11176053_SNP 67440-67441, 67477, 67481, 67500, 67508, 67451, 67526- 67544-67549 67512, 67550- 67543 67559 12:40330692_A_C 67560-67607 67608-67657 67658-67709 rs17444124_REF 12:40330692_A_C 67571, 67573, 67619, 67621, 67669, 67673- rs17444124_SNP 67575-67576, 67623-67625, 67676, 67701, 67594-67595, 67644, 67650, 67703-67706, 67597, 67601, 67652-67654, 67788-67829 67603-67604, 67748-67787 67710-67747 12:40330779_G_A 67830-67877 67878-67925 67926-67976 rs11564203_REF 12:40330779_G_A 67832-67833, 67879, 67881, 67927, 67929, rs11564203_SNP 67837, 67840, 67885, 67896, 67933, 67948- 67849, 67852, 67899, 67901, 67949, 67951, 67854, 67856, 67903, 67920, 67953, 67955, 67860, 67875, 68014-68042 67971, 68043- 67977-68013 68077 12:40331158_G_T 68078-68123 68124-68167 68168-68215 rs7307562_REF 12:40331158_G_T 68078, 68089- 68124-68125, 68168, 68170, rs7307562_SNP 68091, 68099, 68136, 68159, 68181-68182, 68111, 68115, 68164, 68253- 68184, 68212, 68120, 68216- 68281 68282-68316 68252 12:40331402_T_A 68317-68364 68365-68414 68415-68466 rs17520251_REF 12:40331402_T_A 68317-68320, 68365-68368, 68415-68418, rs17520251_SNP 68324, 68326, 68372, 68374, 68430, 68436, 68332, 68334, 68380, 68386, 68443, 68447, 68344, 68350, 68393, 68399, 68450, 68454, 68467-68504 68505-68544 68545-68586 12:40331597_C_CA 68587 , 68588 rs11461395_REF 12:40331769_T_C 68589-68616 68617-68642 68643-68670 rs10506153_REF 12:40331769_T_C 68594, 68598, 68622, 68625, 68648, 68651- rs10506153_SNP 68600, 68605, 68632, 68636, 68652, 68659, 68611, 68671- 68638, 68696- 68663, 68721- 68695 68720 68747 12:40331791_C_G 68748-68758 rs73275761_REF 12:40331791_C_G 68759-68770 rs73275761_SNP 12:40331976_G_A 68771-68818 68819-68868 68869-68920 rs963243_REF 12:40331976_G_A 68778-68779, 68826, 68833- 68874, 68877- rs963243_SNP 68785, 68793, 68834, 68842- 68878, 68885- 68799, 68804- 68843, 68849, 68886, 68894, 68805, 68808- 68854-68855, 68901, 68906- 68809, 68811, 68858-68859, 68907, 68910, 68921-68958 68959-68994 68995-69034 12:40332078_C_T 69035-69080 69081-69130 69131-69182 rs17444145_REF 12:40332078_C_T 69045-69046, 69091-69092, 69135, 69142- rs17444145_SNP 69057, 69065, 69094, 69099, 69143, 69145, 69067, 69069, 69105, 69114, 69150, 69161, 69072-69073, 69116, 69118, 69166, 69170, 69077-69078, 69122, 69127, 69174, 69179, 69183-69220 69221-69260 69261-69302 12:40332126_A_G 69303-69348 69349-69398 69399-69450 rs10878386_REF 12:40332126_A_G 69319, 69332- 69362, 69381- 69412, 69431, rs10878386_SNP 69333, 69335, 69382, 69384, 69434-69435, 69337, 69339, 69388, 69391, 69439, 69443, 69342, 69344- 69393-69395, 69445-69447, 69346, 69451- 69397, 69489- 69449, 69529- 69488 69528 69570 12:40332765_C_G 69571-69572 rs17466521_REF 12:40332765_C_G 69571-69572 rs17466521_SNP 12:40332845_A_T 69573-69604 69605-69630 69631-69658 rs12367542_REF 12:40332845_A_T 69579, 69590- 69609, 69611, 69636, 69638- rs12367542_SNP 69591, 69599- 69620, 69626- 69639, 69648, 69600, 69659- 69627, 69686- 69654, 69707- 69685 69706 69729 12:40332946_T_G 69730-69768 69769-69798 69799-69828 rs17520278_REF 12:40332946_T_G 69768, 69829- 69867-69905 69906-69943 rs17520278_SNP 69866 12:40332960_T_TAA 69738, 69745- 69784, 69790, 69814, 69819, rs35031086_REF 69746, 69748, 69793-69798 69821-69828 69754, 69758- 69768, 69944- 69945 12:40332960_T_TA 69738, 69745- 69784, 69790, 69814, 69819, rs35031086_REF 69746, 69748, 69793-69798 69821-69828 69754, 69758- 69768, 69944- 69945 12:40333796_C_T 69946-69992 69993-70042 70043-70094 rs11834950_REF 12:40333796_C_T 69947-69948, 69993, 69995- 70043, 70046, rs11834950_SNP 69954-69956, 69996, 70002, 70052, 70054- 69965, 69967, 70004-70005, 70055, 70073, 69976, 69982, 70014, 70025, 70076-70077, 69986, 70095- 70031, 70035, 70083, 70087, 70132 70133-70172 70173-70214 12:40333977_T_G 70215-70262 70263-70312 70313-70364 rs11829088_REF 12:40333977_T_G 70222, 70229, 70267, 70271, 70313, 70318, rs11829088_SNP 70232-70233, 70278, 70281- 70322, 70330, 70235, 70239, 70282, 70284, 70333-70334, 70246, 70249, 70288, 70295, 70336, 70347, 70259, 70261, 70298, 70308, 70350, 70360, 70365-70402 70403-70442 70443-70484 12:40334389_C_A 70485-70520 70521-70553 70554-70591 rs4465428_REF 12:40334389_C_A 70502-70505, 70538-70540, 70568, 70573- rs4465428_SNP 70508, 70515, 70542, 70548, 70575, 70577, 70592-70610 70611-70630 70585, 70631- 70655 12:40334496_A_G 70656-70659 rs61007767_REF 12:40334496_A_G 70656-70664 70665-70667 rs61007767_SNP 12:40334626_G_C 70668-70680 70681-70697 70698-70718 rs60776971_REF 12:40334626_G_C 70671, 70719- 70681, 70686, 70699, 70705- rs60776971_SNP 70760 70761-70801 70706, 70802- 70846 12:40335205_C_T 70847-70893 70894-70943 70944-70995 rs2404834_REF 12:40335205_C_T 70856, 70860, 70907, 70911, 70957, 70965- rs2404834_SNP 70862-70863, 70915, 70929- 70966, 70980- 70867, 70881- 70932, 70934, 70981, 70983, 70882, 70884, 70942-70943, 70985, 70993- 70892-70893, 71034-71073 70995, 71074- 70996-71033 71115 12:40335853_G_A 71116-71163 71164-71213 71214-71265 rs7302841_REF 12:40335853_G_A 71118, 71126, 71167, 71175- 71217, 71225- rs7302841_SNP 71140, 71144, 71176, 71190, 71226, 71231, 71148, 71153, 71194, 71198, 71244, 71246, 71156, 71159, 71203, 71209, 71250, 71261, 71161, 71163, 71211, 71213, 71263, 71265, 71266-71303 71304-71343 71344-71385 12:40336661_C_T 71386-71433 71434-71482 71483-71534 rs2404835_REF 12:40336661_C_T 71393, 71395, 71443, 71453, 71491, 71493, rs2404835_SNP 71415-71416, 71462, 71465- 71501, 71504, 71418, 71426, 71466, 71468, 71513, 71516, 71429-71430, 71476, 71480, 71519, 71527, 71432-71433, 71482, 71573- 71531, 71534, 71535-71572 71610 71611-71652 12:40337133_CA_C 71653-71700 71701-71746 71747-71798 rs11356084_REF 12:40337133_CA_C 71654, 71660, 71708, 71711- 71750, 71756, rs11356084_SNP 71663, 71668, 71712, 71715, 71760-71761, 71675, 71681, 71722, 71725, 71766, 71773, 71684, 71688, 71731, 71734, 71776, 71786, 71690, 71799- 71736, 71835- 71788, 71873- 71834 71872 71912 12:40337381_A_C 71913-71916 71917-71919 71920-71924 rs715403_REF 12:40337381_A_C 71913-71916, 71917-71919, 71920-71924, rs715403_SNP 71925-71929 71930 71931-71933 12:40337412_G_A 71934-71978 71979-72010 72011-72050 rs715402_REF 12:40337412_G_A 71934, 71937, 71980, 71989, 72012, 72015, rs715402_SNP 71941, 71949, 71998, 72070- 72035, 72038, 71952, 71965, 72076 72045, 72077- 71969, 72051- 72091 72069 12:40337704_G_C 72092-72123 72124-72149 72150-72177 rs6581667_REF 12:40337704_G_C 72095, 72100, 72127, 72134, 72154, 72158, rs6581667_SNP 72106, 72108, 72136, 72143, 72164, 72171, 72116, 72178- 72148, 72207- 72176, 72228- 72206 72227 72250 12:40337804_G_A 72251-72298 72299-72342 72343-72389 rs6581668_REF 12:40337804_G_A 72253, 72272, 72301, 72308, 72345, 72354, rs6581668_SNP 72274, 72280, 72323, 72325, 72369, 72371, 72282, 72284, 72327-72328, 72373-72374, 72287, 72291, 72335, 72337- 72376, 72382, 72293-72294, 72338, 72428- 72384-72385, 72390-72427 72467 72468-72509 12:40338157_CCCT_C 72510-72556 72557-72606 72607-72658 rs200521371_REF 12:40338157_CCCT_C 72520, 72522, 72559, 72571, 72609, 72627- rs200521371_SNP 72529, 72538, 72578, 72588, 72628, 72638, 72547, 72551, 72597, 72599, 72642, 72648, 72553, 72659- 72603, 72697- 72651, 72737- 72696 72736 72778 12:40338159_C_T 72510-72527, 72557-72576, 72607-72632, rs4767970_REF 72529-72534, 72578-72582, 72634, 72636, 72536, 72538- 72584, 72586, 72638-72649, 72548, 72550- 72588-72606, 72651-72658, 72556, 72661, 72720, 72724, 72745, 72762, 72685, 72779- 72781-72782 72783-72784 72780 12:40338159_C_T 72513, 72520, 72559, 72568, 72609, 72621, rs4767970_SNP 72522, 72529, 72571, 72578, 72627-72628, 72538, 72547, 72588, 72597, 72638, 72642, 72551, 72553, 72599, 72603, 72648, 72651, 72661, 72685- 72720, 72724, 72745, 72762, 72686, 72785- 72732, 72822- 72775, 72861- 72821 72860 72901 12:40338230_C_T 72902-72949 72950-72999 73000-73051 rs4767971_REF 12:40338230_C_T 72902-72903, 72950, 72953- 73004, 73013, rs4767971_SNP 72905-72906, 72954, 72963, 73015-73016, 72914-72915, 72965-72966, 73028, 73033, 72917-72918, 72978, 72986- 73037-73038, 72937, 72946, 72987, 72996, 73043, 73048, 73052-73089 73090-73129 73130-73171 12:40338321_C_T 73172-73209 73210-73235 73236-73265 rs4767972_REF 12:40338321_C_T 73172-73173, 73210-73211, 73236-73238, rs4767972_SNP 73182, 73198, 73215, 73219, 73248, 73258, 73205, 73266- 73229, 73283- 73300-73318 73282 73299 12:40338592_T_C 73319-73341 73342-73345 73346-73351 rs1427271_REF 12:40338592_T_C 73352-73384 73385-73412 73413-73446 rs1427271_SNP 12:40338678_G_C 73447-73487 73488-73526 73527-73567 rs1427272_REF 12:40338678_G_C 73452, 73460, 73606-73644 73645-73685 rs1427272_SNP 73464, 73568- 73605 12:40338928_C_G 73686-73733 73734-73779 73780-73828 rs1427273_REF 12:40338928_C_G 73686, 73690- 73737, 73745, 73783, 73786, rs1427273_SNP 73691, 73699, 73754, 73760, 73792, 73802, 73701, 73711, 73764, 73769, 73808-73809, 73716, 73718, 73867-73906 73813, 73907- 73723, 73733, 73948 73829-73866 12:40339098 C A 73949-73993 73994-74041 74042-74091 rs10735934 REF 12:40339098_C_A 73949, 73951, 73994, 73996, 74042, 74044, rs10735934_SNP 73954, 73959, 73999, 74004- 74047, 74053, 73968, 73972, 74005, 74014, 74062, 74071, 73980-73981, 74016, 74028, 74076, 74078, 74092-74128 74129-74165 74166-74207 12:40339280_A_G 74208-74225 74226-74241 74242-74261 rs10732751_REF 12:40339280_A_G 74213, 74218, 74229, 74234, 74247, 74252, rs10732751_SNP 74221, 74224- 74237, 74240- 74255, 74259, 74225, 74262- 74241, 74285- 74261, 74300- 74284 74299 74318 12:40339762_T_G 74319-74337 74338-74348 74349-74363 rs12306060_REF 12:40339762_T_G 74319-74320, 74338, 74343, 74354-74355, rs12306060_SNP 74323, 74328, 74383-74399 74357, 74359, 74364-74382 74400-74420 12:40340781_T_C 74421-74465 74466-74505 74506-74548 rs1365763_REF 12:40340781_T_C 74427, 74429, 74468, 74470, 74506, 74510, rs1365763_SNP 74434-74435, 74475, 74479, 74512, 74517, 74440, 74446- 74485-74486, 74521, 74527, 74447, 74459, 74498, 74502, 74540, 74544, 74463, 74465, 74505, 74587- 74547, 74625- 74549-74586 74624 74666 12:40341114_C_A 74667-74714 74715-74764 74765-74816 rs7963697_REF 12:40341114_C_A 74667-74668, 74715-74716, 74765, 74786- rs7963697_SNP 74670, 74674, 74736-74737, 74788, 74790, 74688, 74690, 74739, 74741, 74801, 74807, 74692, 74706, 74750, 74756, 74809-74810, 74708-74709, 74758-74759, 74814, 74895- 74817-74854 74855-74894 74936 12:40341174_TG_T 74937-74983 74984-75033 75034-75085 rs11316380_REF 12:40341174_TG_T 74940, 74946, 74988, 74995, 75045, 75056, rs11316380_SNP 74948, 74962- 75006, 75012, 75064, 75066, 74963, 74965, 75014, 75017, 75068, 75073, 74968, 74978, 75022, 75028, 75079, 75081, 74981, 75086- 75031, 75122- 75083, 75160- 75121 75159 75199 12:40341367_G_C 75200-75245 75246-75291 75292-75340 rs7963086_REF 12:40341367_G_C 75200, 75202, 75246, 75248, 75292, 75294, rs7963086_SNP 75204, 75208, 75250, 75254, 75296, 75305, 75213, 75218, 75266, 75282, 75313, 75317- 75224, 75236, 75379-75418 75318, 75419- 75341-75378 75460 12:40341862_A_G 75461-75466 75467-75471 75472-75477 rs17491536_REF 12:40341862_A_G 75461, 75463- 75467-75471, 75472-75473, rs17491536_SNP 75466, 75478- 75487-75489 75475-75477, 75486 75490-75494 12:40341962_T_C 75495-75541 75542-75585 75586-75634 rs7956787_REF 12:40341962_T_C 75495-75496, 75548, 75553, 75593, 75599, rs7956787_SNP 75504-75505, 75557, 75559, 75603, 75605, 75510, 75514, 75564, 75576- 75610, 75616, 75516, 75521, 75577, 75673- 75623-75624, 75534, 75538, 75712 75713-75754 75635-75672 12:40342002_T_G 75755-75802 75803-75846 75847-75898 rs7956898_REF 12:40342002_T_G 75759, 75765, 75806, 75809, 75857, 75865, rs7956898_SNP 75767, 75769, 75817-75818, 75867, 75871, 75773, 75777, 75822, 75828, 75877, 75879, 75783, 75785, 75830, 75832, 75881, 75883, 75787, 75793, 75838, 75841, 75888, 75892, 75899-75936 75937-75976 75977-76018 12:40342137_G_A 76019-76066 76067-76116 76117-76168 rs10506155_REF 12:40342137_G_A 76019, 76021, 76069, 76075, 76119, 76125, rs10506155_SNP 76027, 76031, 76078, 76080, 76128, 76130- 76041, 76049- 76090, 76099, 76131, 76153, 76050, 76053, 76102, 76107, 76158, 76163, 76058, 76064, 76112, 76114, 76165, 76168, 76169-76206 76207-76246 76247-76288 12:40342208_A_G 76289-76336 76337-76386 76387-76438 rs7954061_REF 12:40342208_A_G 76289-76290, 76337-76338, 76387-76388, rs7954061_SNP 76295, 76300- 76348-76349, 76390, 76399- 76301, 76304, 76352, 76358, 76400, 76403, 76315, 76324, 76361, 76374, 76409, 76412, 76326, 76330, 76376, 76380, 76427, 76430, 76439-76476 76477-76516 76517-76558 12:40342241_T_A 76559-76606 76607-76656 76657-76708 rs7957151_REF 12:40342241_T_A 76559, 76561, 76607, 76609, 76657, 76659, rs7957151_SNP 76564, 76566- 76612, 76614- 76664, 76673, 76567, 76572, 76615, 76623, 76679-76680, 76575, 76584, 76632, 76637, 76684, 76689, 76589-76590, 76646, 76649, 76698, 76701, 76709-76746 76747-76786 76787-76828 12:40342447_C_T 76829-76876 76877-76925 76926-76977 rs919174_REF 12:40342447_C_T 76831-76832, 76879, 76881, 76926, 76929, rs919174_SNP 76838, 76842, 76887, 76891, 76937, 76941, 76845-76846, 76895, 76905, 76945, 76955, 76856, 76859, 76908-76909, 76958-76960, 76862-76863, 76912, 77016- 76964, 77051- 76978-77015 77050 77089 12:40342479_C_CTT 77090-77101 77102-77113 77114-77127 rs3065921_REF 12:40342479_C_CT 77090-77101 77102-77113 77114-77127 rs3065921_REF 12:40342603_C_T 77128-77165 77166-77201 77202-77239 rs919175_REF 12:40342603_C_T 77138, 77153- 77176-77177, 77212-77213, rs919175_SNP 77154, 77161- 77191-77192, 77228, 77235- 77162, 77240- 77198, 77269- 77236, 77298- 77268 77297 77328 12:40342980_C_T 77329-77348 77349-77366 77367-77386 rs1035812_REF 12:40342980_C_T 77333, 77338, 77357, 77359, 77374, 77377, rs1035812_SNP 77341, 77346, 77364-77366, 77379, 77385- 77348, 77387- 77398-77408 77386, 77409- 77397 77421 12:40343200_G_A 77422-77441 77442-77455 77456-77472 rs1365764_REF 12:40343200_G_A 77426, 77428, 77444-77445, 77459-77462, rs1365764_SNP 77431, 77437, 77447, 77452, 77464, 77469, 77440, 77473- 77454, 77480- 77485-77491 77479 77484 12:40343257_C_CTCAG 77492-77538 77539-77585 77586-77635 rs397724721_REF 12:40343257_C_CTCAG 77492, 77503, 77548, 77550, 77596, 77598- rs397724721_SNP 77519, 77522, 77567, 77570- 77599, 77613, 77536, 77538, 77571, 77585, 77615, 77619, 77636-77660 77661-77687 77688-77716 12:40343273_C_T 77492-77494, 77539-77541, 77586-77588, rs17461964_REF 77496-77498, 77543-77545, 77591-77593, 77504, 77507- 77551, 77555- 77599, 77603- 77508, 77511, 77557, 77559, 77605, 77607, 77523-77524, 77571-77572, 77616, 77620- 77526, 77534- 77581-77582, 77621, 77623, 77535, 77717- 77732-77740 77625, 77631- 77731 77632, 77741- 77752 12:40343273_C_T 77493, 77497, 77540, 77544, 77587, 77592, rs17461964_SNP 77508, 77511, 77556-77557, 77605, 77607, 77535, 77721, 77559, 77732, 77625, 77742- 77723-77724, 77735-77736, 77743, 77747, 77726, 77731, 77738, 77740, 77749, 77752, 77753-77763 77764-77772 77773-77784 12:40343313_G_T 77785-77830 77831-77880 77881-77932 rs10784518_REF 12:40343313_G_T 77785-77786, 77831-77832, 77881, 77889, rs10784518_SNP 77798, 77804, 77845, 77851- 77896, 77902- 77811, 77814, 77852, 77859, 77904, 77911, 77821, 77825- 77864, 77870, 77916, 77922, 77826, 77933- 77875-77876, 77927, 78011- 77970 77971-78010 78052 12:40343381_G_A 78053-78073 78074-78082 78083-78097 rs4768231_REF 12:40343381_G_A 78056, 78058, 78105-78110 78090, 78111- rs4768231_SNP 78069-78070, 78118 78098-78104 12:40343438_C_T 78119-78164 78165-78211 78212-78262 rs4768232_REF 12:40343438_C_T 78125, 78128, 78171, 78173, 78218, 78220, rs4768232_SNP 78133, 78140- 78176, 78181, 78223, 78228, 78141, 78148, 78189, 78196, 78230, 78237, 78155, 78158- 78203, 78299- 78245, 78250, 78159, 78263- 78333 78253, 78334- 78298 78374 12:40344001_A_G 78375-78402 78403-78408 78409-78418 rs3943893_REF 12:40344001_A_G 78419-78455 78456-78487 78488-78526 rs3943893_SNP 12:40344030_T_C 78527-78536 rs2162471_REF 12:40344030_T_C 78530, 78533, 78560-78570 78571-78587 rs2162471_SNP 78537-78559 12:40344106_C_T 78588-78609 78610-78631 78632-78653 rs1035813_REF 12:40344106_C_T 78593, 78596, 78618-78619, 78640, 78643, rs1035813_SNP 78601-78602, 78623-78624, 78645-78646, 78604, 78606, 78626, 78628, 78650-78651, 78608, 78654- 78630, 78669- 78653, 78682- 78668 78681 78696 12:40344206_C_T 78697-78731 78732-78760 78761-78796 rs10459264_REF 12:40344206_C_T 78697-78699, 78732-78733, 78761, 78770, rs10459264_SNP 78703, 78712- 78741, 78744, 78773-78774, 78713, 78718- 78719, 78797- 78747, 78751, 78778-78779, 78817 78818-78838 78784-78785, 78839-78861 12:40344501_G_A 78862-78909 78910-78959 78960-79011 rs10459265_REF 12:40344501_G_A 78878, 78880, 78916, 78925, 78961, 78964, rs10459265_SNP 78885-78886, 78928, 78935- 78968, 78977, 78894-78895, 78936, 78944- 78980, 78988, 78897-78898, 78945, 78948, 78996, 79000, 78900, 78905, 78950, 78955, 79002, 79007, 79012-79049 79050-79085 79086-79127 12:40345691_G_A 79128-79165 79166-79202 79203-79241 rs11564147_REF 12:40345691_G_A 79131, 79140, 79169, 79178, 79206, 79209, rs11564147_SNP 79145, 79153, 79183, 79190, 79221, 79229, 79164, 79242- 79273-79303 79304-79336 79272 12:40345826_TA_T 79337-79340 rs10715759_REF 12:40345826_TA_T 79337, 79339, rs10715759_SNP 79341-79346 12:40345925_C_T 79347-79385 79386-79422 79423-79463 rs7296657_REF 12:40345925_C_T 79368, 79380, 79417, 79421, 79461, 79463, rs7296657_SNP 79384, 79464- 79497-79519 79520-79548 79496 12:40346050_A_T 79549-79562 79563-79576 79577-79592 rs7312497_REF 12:40346050_A_T 79554, 79556, 79569, 79571, 79581, 79584, rs7312497_SNP 79558, 79560, 79573, 79575- 79589, 79591- 79562, 79593- 79576, 79602- 79592, 79611- 79601 79610 79621 12:40346187_CT_C 79622-79639 79640-79657 79658-79675 rs10715758_REF 12:40346187_CT_C 79634-79635 79646, 79652 79662, 79664 rs10715758_SNP 12:40346563_G_T 79676-79723 79724-79773 79774-79825 rs10784522_REF 12:40346563_G_T 79683, 79692, 79731, 79740, 79781, 79790, rs10784522_SNP 79697, 79703- 79745, 79751, 79795, 79801, 79704, 79708, 79757, 79761, 79807, 79811, 79712, 79718- 79767-79768, 79817, 79822, 79719, 79723, 79771, 79773, 79824-79825, 79826-79863 79864-79903 79904-79945 12:40347026_AT_A 79946-79954 79955 79956-79957 rs796827140_REF 12:40347026_AT_A 79950 rs796827140_SNP 12:40347033_T_G 79950, 79953 rs74467833_REF 12:40347033_T_G 79950, 79953, 79966-79967 rs74467833_SNP 79958-79965 12:40347145_A_G 79968-79970 79971-79973 79974-79977 rs7131694_REF 12:40347145_A_G 79968-79970 79971-79973 79974-79977 rs7131694_SNP 12:40347815_T_C 79978-79979 79980-79983 79984-79989 rs12311273_REF 12:40347815_T_C 79978, 79990- 79980, 79982, 79985, 79987- rs12311273_SNP 79991 79992-79994 79988, 79995- 80000 12:40347876_T_C 80001-80005 80006-80014 80015-80027 rs58911468_REF 12:40347876_T_C 80001-80003, 80007-80009, 80016, 80020- rs58911468_SNP 80028-80042 80013, 80043- 80021, 80023, 80060 80026, 80061- 80081 12:40347956_T_TAA 80082-80084 80085-80089 80090-80096 rs35847030_REF 12:40347956_T_TA 80082-80084 80085-80089 80090-80096 rs35847030_REF 12:40348019_A_G 80097-80124 80125-80148 80149-80178 rs1365765_REF 12:40348019_A_G 80101-80103, 80129-80131, 80155-80157, rs1365765_SNP 80105-80107, 80134, 80136- 80160, 80162, 80109-80110, 80137, 80139- 80165, 80168- 80118, 80120, 80140, 80145- 80169, 80174- 80179-80210 80146, 80211- 80175, 80235- 80234 80268 12:40348202_AT_A 80269-80278 80279-80287 80288-80299 rs66513412_REF 12:40348202_AT_A 80270, 80277- 80280, 80287, 80289-80290, rs66513412_SNP 80278, 80300- 80306-80311 80293, 80312- 80305 80319 12:40348203_T_C 80270-80272, 80280, 80282- 80289-80293, rs2896977_REF 80274-80278 80287 80295-80299 12:40348203_T_C 80270, 80277- 80280, 80287, 80289-80290, rs2896977_SNP 80278, 80320- 80327-80333 80293, 80334- 80326 80342 12:40348204_A_C 80270-80272, 80280, 80282, 80289, 80291, rs199606989_REF 80275-80278, 80284-80287 80293, 80295- 80343 80299 12:40348204_A_C 80270, 80275, 80280, 80286- 80289, 80293, rs_199606989_SNP 80277-80278, 80287, 80349- 80299, 80354- 80343-80348 80353 80360 12:40348271_C_T 80361-80364 , , rs1365766_REF 12:40348452_G_A 80365-80384 80385-80406 80407-80430 rs10878405_REF 12:40348452_G_A 80374, 80377, 80396-80397, 80419-80421, rs10878405_SNP 80379, 80381, 80400, 80402, 80426, 80429, 80383, 80431- 80405, 80446- 80461-80477 80445 80460 12:40348561_G_A 80478-80488 80489-80492 80493-80500 rs11176143_REF 12:40348561_G_A 80479, 80482, 80491-80492 80497, 80499- rs11176143_SNP 80486-80487 80500 12:40348887_T_C 80501-80531 80532-80562 80563-80600 rs7294958_REF 12:40348887_T_C 80505, 80509- 80537, 80540- 80567, 80569, rs7294958_SNP 80510, 80512, 80541, 80543, 80573-80574, 80518-80519, 80546, 80549- 80577, 80580, 80522, 80524- 80550, 80554, 80585, 80590, 80525, 80528, 80556, 80559, 80592, 80596, 80601-80634 80635-80666 80667-80704 12:40348899_C_T 80501-80506, 80532-80542, 80563-80566, rs61915610_REF 80508-80511, 80544, 80551- 80568-80570, 80513, 80520- 80555, 80557- 80572-80574, 80523, 80526- 80562, 80724- 80576, 80578, 80531, 80705- 80741 80581, 80587- 80723 80591, 80593- 80600, 80742- 80759 12:40348899_C_T 80520, 80526, 80557, 80561, 80581, 80598, rs61915610_SNP 80530, 80760- 80797-80833 80600, 80834- 80796 80872 12:40349074_A_T 80873-80890 80891-80892 80893-80898 rs7294952_REF 12:40349074_A_T 80873, 80878, 80912-80913 80914-80919 rs7294952_SNP 80882, 80887, 80890, 80899- 80911 12:40349558_GGCTGGAGT_G 80920 rs59096461_REF 12:40349690_A_AT 80921-80926 80927-80933 80934-80943 rs59980086_REF 12:40349986_C_T 80944-80991 80992-81037 81038-81086 rs4768233_REF 12:40349986_C_T 80946-80947, 81005-81006, 81041, 81052- rs4768233_SNP 80955, 80959, 81019, 81026, 81053, 81065, 80972, 80975, 81031, 81034, 81075, 81080, 80980, 80985, 81123-81148 81083, 81149- 80988, 80991, 81180 81087-81122 12:40350028_G_A 81181-81228 81229-81277 81278-81329 rs4768234_REF 12:40350028_G_A 81182, 81184, 81230, 81232, 81279, 81281, rs4768234_SNP 81188, 81194, 81236, 81246- 81287, 81291, 81198-81199, 81247, 81256, 81297-81298, 81212, 81216, 81261, 81271, 81305, 81308, 81222, 81227, 81276, 81366- 81323, 81328, 81330-81365 81395 81396-81435 12:40350284_A_T 81436-81447 81448-81459 81460-81474 rs4768235_REF 12:40350284_A_T 81436-81437, 81448-81450, 81460-81462, rs4768235_SNP 81439, 81441- 81452-81453, 81464, 81466- 81444, 81446- 81455, 81457- 81467, 81469, 81447, 81475- 81459, 81478- 81471, 81473- 81477 81480 81474, 81481- 81485 12:40350343_C_A 81486-81533 81534-81583 81584-81635 rs7314455_REF 12:40350343_C_A 81489, 81492, 81537, 81540- 81590-81591, rs7314455_SNP 81495, 81500- 81541, 81544, 81594, 81602, 81501, 81509- 81552, 81559- 81609-81611, 81510, 81513, 81560, 81563, 81616, 81618, 81515, 81517, 81565, 81567, 81628, 81714- 81636-81673 81674-81713 81755 12:40350381_G_A 81756-81803 81804-81853 81854-81905 rs7313525_REF 12:40350381_G_A 81758-81759, 81806, 81815, 81856, 81874, rs7313525_SNP 81767, 81776, 81824, 81828, 81878, 81883, 81779, 81786, 81833, 81836, 81886, 81894, 81794-81795, 81844-81845, 81896, 81898, 81801, 81803, 81851, 81853, 81903, 81905, 81906-81943 81944-81983 81984-82025 12:40350592_G_T 82026-82046 82047-82066 82067-82088 rs7313895_REF 12:40350592_G_T 82031-82032, 82053, 82059- 82071, 82074, rs7313895_SNP 82040, 82044, 82060, 82064, 82082, 82086, 82046, 82089- 82066, 82102- 82088, 82113- 82101 82112 82125 12:40350689_C_T 82126-82137 82138-82142 82143-82149 rs7314863_REF 12:40350689_C_T 82126-82128, rs7314863_SNP 82150-82152 12:40350973_A_C 82153-82200 82201-82250 82251-82302 rs7303525_REF 12:40350973_A_C 82158, 82164, 82212, 82216, 82261, 82263, rs7303525_SNP 82169, 82184, 82231, 82234, 82267, 82277, 82186, 82191, 82236, 82241, 82283, 82288, 82194, 82198- 82244, 82248- 82293, 82300- 82200, 82303- 82250, 82341- 82302, 82381- 82340 82380 82422 12:40351006_G_A 82423-82469 82470-82519 82520-82571 rs7132187_REF 12:40351006_G__A 82425, 82439- 82472-82473, 82522-82523, rs7132187_SNP 82441, 82443, 82489, 82491, 82539, 82541, 82448, 82458, 82496-82497, 82547, 82550, 82467, 82469, 82505, 82508, 82556, 82568- 82572-82609 82517, 82519, 82569, 82571, 82610-82649 82650-82691 12:40351379_T_C 82692 82693 rs7307310_REF 12:40351379_T_C 82692 82693 rs7307310_SNP 12:40352036_T_C 82694-82740 82741-82790 82791-82842 rs11176153_REF 12:40352036_T_C 82694-82695, 82741-82742, 82791, 82800, rs11176153_SNP 82697, 82703, 82750, 82753, 82803, 82806, 82716, 82719, 82756, 82765, 82815, 82818, 82723-82724, 82768, 82772, 82822-82824, 82728, 82737, 82777, 82786, 82829, 82921- 82843-82880 82881-82920 82962 12:40352100_G_C 82963-83009 83010-83059 83060-83111 rs11176154_REF 12:40352100_G_C 82980, 82982- 83027, 83029- 83068, 83078, rs11176154_SNP 82984, 82994, 83031, 83033, 83080-83083, 83001, 83004- 83041, 83050- 83085, 83093, 83005, 83009, 83051, 83055, 83102-83103, 83112-83149 83059, 83150- 83190-83231 83189 12:40352112_T_C 82965, 82968- 83012, 83015- 83062, 83065- rs7311247_REF 82971, 82973, 83018, 83020, 83067, 83069, 82977-82982, 83024-83029, 83071, 83075- 82985, 82989- 83032-83033, 83081, 83084- 82992, 82995- 83035, 83037- 83085, 83087, 82996, 82999, 83040, 83044- 83089-83092, 83004, 83006, 83045, 83048, 83096-83097, 83009, 83232- 83050, 83054, 83100, 83102, 83255 83056, 83059, 83105-83106, 83256-83279 83108, 83111, 83280-83303 12:40352112_T_C 82965, 82971, 83012, 83018, 83062, 83071, rs7311247_SNP 82973, 82995, 83020, 83035, 83087, 83100, 82999, 83232- 83048, 83256- 83105, 83280- 83233, 83237, 83257, 83261, 83281, 83286, 83245, 83247, 83269, 83275, 83293, 83299, 83304-83341 83342-83381 83382-83423 12:40352153_C_T 83424-83470 83471-83514 83515-83563 rs7137437_REF 12:40352153_C_T 83428, 83430, 83475, 83485, 83521, 83530, rs7137437_SNP 83439, 83444- 83487, 83491- 83532, 83536- 83445, 83448, 83492, 83500, 83538, 83540, 83451, 83454, 83506, 83595- 83548, 83554, 83461, 83564- 83617 83618-83646 83594 12:40352235_AG_A 83647-83694 83695-83742 83743-83793 rs75439407_REF 12:40352235_AG_A 83647, 83652, 83695, 83700, 83743, 83748, rs75439407_SNP 83657, 83669, 83705, 83721, 83753, 83770- 83675-83676, 83724, 83728, 83771, 83775, 83680, 83690, 83692, 83794- 83738, 83828- 83778, 83791, 83827 83861 83862-83899 12:40352271_T_C 83900-83947 83948-83997 83998-84049 rs73107108_REF 12:40352271_T_C 83900, 83902, 83948, 83956, 84006, 84008, rs73107108_SNP 83908, 83910, 83958, 83963, 84013, 84015- 83915, 83919, 83965, 83968, 84016, 84028, 83928, 83930, 83977, 83979, 84030, 84033, 83935-83936, 83984, 83994, 84036, 84046, 84050-84087 84088-84127 84128-84169 12:40352469_C_CTTTTTTTTT 84170-84183 84184-84190 84191-84201 TTTTTT rs1307806388_REF 12:40352469_C_CTTTTTTTTT 84170-84183 84184-84190 84191-84201 TTTTT rs1307806388_REF 12:40352469_C_CTTTTTTTTT 84170-84183 84184-84190 84191-84201 TTTT rs1307806388_REF 12:40352469_C_CT 84170-84183 84184-84190 84191-84201 rs1307806388_REF 12:40352707_T_G 84202-84244 84245-84294 84295-84346 rs10878409_SNP 12:40352874_C_T 84347-84357 84358-84368 84369-84381 rs10878410_SNP 12:40352927_G_C 84382-84403 84404-84427 84428-84453 rs138382676_REF 12:40352927_G_C 84385-84386, 84407-84409, 84431-84433, rs138382676_SNP 84393, 84399, 84423, 84483- 84443, 84517- 84454-84482 84516 84554 12:40353080_G_A A , 84555 rs11176161_SNP 12:40353111_T_C 84556-84588 84589-84623 84624-84660 rs10878412_REF 12:40353111_T_C 84557-84558, 84591, 84605- 84626, 84642, rs10878412_SNP 84571-84572, 84606, 84610, 84646, 84653, 84576, 84661- 84621, 84683- 84658, 84688- 84682 84687 84701 12:40353158_T_C , 84702 84703-84705 rs10878413_REF 12:40353186_C_T 84706-84716 84717-84725 84726-84736 rs183544696_REF 12:40353186_C_T 84737-84776 84777-84813 84814-84853 rs183544696_SNP 12:40353191_C_T 84706-84716 84717-84725 84726-84736 rs189538849_REF 12:40353191_C_T 84854-84886 84887-84913 84914-84943 rs189538849_SNP 12:40353217_C_T 84944-84951 84952-84961 84962-84975 rs11610569_SNP 12:40353226_C_T 84976-84988 84989-85003 85004-85020 rs142726158_REF 12:40353226_C_T 84979-84981, 84991, 84993- 85006, 85008- rs142726158_SNP 84983, 84985, 84995, 84999, 85009, 85014- 85021-85038 85039-85058 85015, 85059- 85080 12:40353242_CAAA_C 84976-84988, 84989-85003, 85004-85020, rs1194726090_REF 85081-85105 85106-85132 85133-85159 12:40353242_CAAA_C 84977-84978, 84992, 84996, 85011, 85016- rs1194726090_SNP 84986, 85160- 85000, 85189- 85017, 85213- 85188 85212 85239 12:40353243_A_G 84976-84988, 84989-85003, 85004-85020, rs576444406_REF 85081-85105 85106-85132 85133-85159, 85240 12:40353243_A_G 84977-84978, 84992, 84996, 85011, 85016- rs576444406_SNP 84982, 84986, 85000-85001, 85018, 85156, 85103, 85174, 85116, 85191, 85220, 85311- 85241-85274 85275-85310 85348 12:40353244_A_G 84976-84986, 84989-85001, 85004-85018, rs185723137_REF 84988, 85081- 85003, 85106- 85020, 85133- 85105 85132 85159, 85240, 85349 12:40353246_GGGGCTCCTCA 84976-84981, 84989-84995, 85004-85011, CTTCTCAGACGGGGCGGCT 84984-84985, 84998-85000, 85013-85016, GCCGGGCA_G 85081-85105 85106-85132 85133-85159, rs1179215763_REF 85240, 85349 12:40353246_GGGGCTCCTCA 84976, 84984, 84989-84990, 85004-85005, CTTCTCAGACGGGGCGGCT 85091, 85100- 84998, 85115, 85007, 85013, GCCGGGCA_G 85101, 85350- 85128, 85390- 85142, 85432- rs1179215763_SNP 85389 85431 85475 12:40353266_C_T , , 85240, 85349 rs71449759_REF 12:40353266_C_T 85476-85478 85479-85483 85349, 85484- rs71449759_SNP 85489 12:40353289_A_G 85490-85494 85495-85501 85502-85510 rs61915613_REF 12:40353344_A_G 85511-85525 85526-85542 85543-85561 rs140722234_REF 12:40353344_A_G 85513-85514, 85528-85529, 85545-85546, rs140722234_SNP 85520, 85523- 85538-85540, 85556-85557, 85524, 85562- 85572-85580 85559, 85581- 85571 85592 12:40353383_C_T 85593-85597 85598-85604 85605-85613 rs191099092_REF 12:40353383_C_T 85593-85594, 85598-85599, 85605-85606, rs191099092_SNP 85597, 85614- 85603-85604, 85611-85613, 85621 85622-85632 85633-85646 12:40353440_C_A 85647-85686 85687-85726 85727-85770 rs113693842_REF 12:40353440_C_A 85655, 85683, 85695, 85700, 85735, 85740- rs113693842_SNP 85771-85775 85724, 85776- 85741, 85768, 85786 85787-85803 12:40353467_C_T 85804-85809 85810-85818 85819-85829 rs112767992_REF 12:40353467_C_T 85804-85805, 85810-85811, 85820, 85823, rs112767992_SNP 85807, 85809, 85814, 85816, 85825, 85827, 85830-85835 85818, 85836- 85829, 85845- 85844 85857 12:40353515_AG_A 85858-85884 85885-85908 85909-85939 rs34531686_REF 12:40353632_C_T 85940 85941-85943 85944-85948 rs12368227_REF 12:40353632_C_T 85940 85941, 85943, 85944-85945, rs12368227_SNP 85949 85948, 85950- 85951 12:40353660_C_A 85952-85986 85987-86022 86023-86061 rs12368229_REF 12:40353660_C_A 85953, 85960, 85994, 85996, 86031, 86038- rs12368229_SNP 85965, 85968, 86001, 86004, 86039, 86042, 85983, 86062- 86019, 86097- 86058, 86134- 86096 86133 86173 12:40353732_G_C , 86174 86175-86176 rs3930031_SNP 12:40354127_C_T 86177-86221 86222-86264 86265-86312 rs7308626_REF 12:40354127_C_T 86189, 86193, 86233, 86238, 86273, 86280, rs7308626_SNP 86195, 86201, 86244, 86248, 86285-86286, 86205, 86211, 86254, 86259, 86292, 86296, 86213, 86313- 86346-86370 86302, 86307, 86345 86371-86401 12:40354593_G_A 86402-86423 86424-86430 86431-86444 rs890575_REF 12:40354593_G_A 86409-86410, 86425, 86427, 86435, 86438, rs890575_SNP 86412, 86419- 86448-86450 86442, 86451- 86420, 86445- 86455 86447 12:40354745_A_G 86456-86503 86504-86553 86554-86605 rs10748040_REF 12:40354745_A_G 86459, 86462, 86507, 86510, 86557, 86560, rs10748040_SNP 86467-86468, 86515-86516, 86565, 86572, 86476, 86478, 86522, 86525, 86575-86576, 86482, 86484, 86527, 86533- 86584-86585, 86489, 86492, 86534, 86539, 86590, 86592, 86606-86643 86644-86683 86684-86725 12:40354835_AAAAAAAAAA 86726-86727 TATAT_A rs61230125_REF 12:40354835_AAAAAAAAAA 86726-86727 TATAT_A rs61230125_SNP 12:40354862_A_C 86728-86743 86744-86757 86758-86773 rs11176165_REF 12:40354862_A_C 86728, 86730, 86744-86745, 86758-86759, rs11176165_SNP 86735-86736, 86749, 86752, 86766, 86768, 86738, 86774- 86754, 86789- 86773, 86802- 86788 86801 86816 12:40355348_G_A 86817-86864 86865-86914 86915-86966 rs7976715_REF 12:40355348_G_A 86817-86818, 86865-86867, 86915-86916, rs7976715_SNP 86820-86821, 86870, 86872, 86918, 86921, 86823, 86830, 86879, 86887, 86923, 86928, 86838, 86850, 86898, 86900, 86931, 86939, 86853, 86863, 86903, 87005- 86950, 86955, 86967-87004 87044 87045-87086 12:40355370_G_A 86823, 86837, 86872, 86874, 86919, 86923, rs7976724_REF 86843, 86853, 86886, 86892, 86925, 86928, 87087-87130 86898, 86903, 86938, 86944, 87131-87174 86950, 86955, 87175-87218 12:40355370_G_A 86837, 86843, 86874, 86886, 86919, 86925, rs7976724_SNP 87095, 87101, 86892, 87145, 86938, 86944, 87105, 87109, 87149, 87153, 87192, 87196- 87112, 87124, 87156, 87169, 87197, 87200, 87126, 87130, 87173-87174, 87213, 87218, 87219-87256 87257-87296 87297-87338 12:40355395_A_G 87339-87385 87386-87435 87193, 87197, rs7963987_REF 87436-87485 12:40355395_A_G 87342, 87344, 87389, 87394, 87193, 87439, rs7963987_SNP 87347, 87351, 87398, 87401, 87444, 87448, 87354, 87362, 87410, 87422, 87458, 87461, 87373, 87375- 87424-87426, 87472, 87474- 87376, 87381, 87431, 87524- 87476, 87564- 87486-87523 87563 87605 12:40355416_G_C 87341-87342, 87388-87389, 87437-87439, rs2162472_REF 87351, 87366, 87397-87398, 87447-87448, 87372, 87376, 87414, 87420, 87458, 87464, 87606-87629 87424, 87426, 87470, 87474, 87630-87654 87476, 87655- 87679 12:40355416_G_C 87341, 87366, 87388, 87397, 87437-87438, rs2162472_SNP 87372, 87609, 87414, 87420, 87447, 87464, 87623, 87680- 87634, 87714- 87470, 87752- 87713 87751 87792 12:40355505_A_ATCCCTCCC 87793-87807 87808-87824 87825-87843 TCCCTCCC rs767501990_REF 12:40355505_A_ATCCCTCCC 87793-87807 87808-87824 87825-87843 TCCC rs767501990_REF 12:40355505_A_ATCCCTCCC 87793-87807 87808-87824 87825-87843 rs767501990_REF 12:40355505_A_ATCCC 87793-87807 87808-87824 87825-87843 rs767501990_REF 12:40355543_T_TCCTTC 87844-87845 87846-87849 87850-87855 rs552326490_REF 12:40355543_T_TC 87844-87845 87846-87849 87850-87855 rs552326490_REF 12:40355544_CTT_C 87844-87845 87846-87849 87850-87855 rs1365381164_REF 12:40355546_T_TC 87844-87845 87846-87849 87850-87855 rs760911090_REF 12:40355546_T_TC 87856 87857 87858-87860 rs760911090_SNP 12:40355549_T_TC 87844-87845 87846-87849 87850-87855 rs754028820_REF 12:40356669_G_A 87861-87908 87909-87958 87959-88010 rs11564173_REF 12:40356669_G_A 87872, 87874, 87918, 87921, 87961, 87969, rs11564173_SNP 87883, 87886, 87927, 87933, 87972, 87978- 87891, 87896, 87936, 87941, 87979, 87985, 87900, 87906- 87946, 87950, 87988, 87993, 87908, 88011- 87957-87958, 88002, 88010, 88048 88049-88088 88089-88130 12:40356747_T_A 88131-88142 88143-88148 88149-88158 rs7306545_REF 12:40356747_T_A 88135-88136, 88147-88148, 88152-88153, rs7306545_SNP 88140-88142, 88166-88169 88156-88157, 88159-88165 88170-88175 12:40357018_A_G 88176 88177 rs7306684_REF 12:40357018_A_G 88178-88216 88217-88248 88249-88284 rs7306684_SNP 12:40357164_C_T 88285 rs112259589_REF 12:40357164_C_T 88286-88300 88301-88319 88285, 88320- rs112259589_SNP 88341 12:40357182_A_G 88342-88358 88359-88375 88285, 88376- rs73277531_REF 88399 12:40357182_A_G 88400-88436 88437-88475 88383, 88476- rs73277531_SNP 88517 12:40357363_CTT_C 88518-88534 88535-88550 88551-88568 rs77689380_REF 12:40357363_CTT_C 88526, 88531, 88542-88543 88560-88561, rs77689380_SNP 88569 88566, 88570 12:40357725_G_A 88571-88606 88607-88643 88644-88683 rs73277533_REF 12:40357725_G_A 88571-88572, 88607, 88621, 88659, 88666, rs73277533_SNP 88590, 88600, 88628, 88718- 88675, 88737- 88684-88717 88736 88759 12:40357968_A_G 88760-88798 88799-88839 88840-88882 rs17462055_REF 12:40357968_A_G 88794-88798, 88835-88839, 88850, 88879- rs17462055_SNP 88883-88919 88920-88960 88882, 88961- 89005 12:40358092_C_CT 89006-89013 89014-89021 89022-89031 rs758880647_REF 12:40358177_T_C 89032-89073 89074-89102 89103-89137 rs10784528_REF 12:40358177_T_C 89042, 89047, 89078, 89083, 89104, 89108, rs10784528_SNP 89053, 89058, 89090, 89098, 89113, 89121, 89063, 89068, 89175-89211 89127, 89131, 89071, 89138- 89212-89250 89174 12:40358735_AT_A 89251-89260 89261-89266 89267-89274 rs796949072_REF 12:40358801_A_T 89275 89276-89277 rs10784532_REF 12:40358801_A_T 89278-89285 89275, 89286- 89276-89277, rs_10784532_SNP 89292 89293-89303 12:40359018_TTTG_T 89304-89333 89334-89349 89350-89371 rs145580704_REF 12:40359018_TTTG_T 89317, 89322, 89344, 89346, 89367, 89369- rs145580704_SNP 89326, 89330, 89348-89349, 89371, 89376- 89372-89374 89375 89377 12:40359242_C_CT 89378-89396 89397-89415 89416-89434 rs386376254_REF 12:40359242_C_CT 89389 89410 89432 rs386376254_SNP 12:40359972_C_T 89435-89446 89447-89457 89458-89470 rs11836288_REF 12:40359972_C_T 89435, 89471- rs11836288_SNP 89479 12:40360506_T_C 89480-89527 89528-89577 89578-89629 rs11564146_REF 12:40360506_T_C 89482, 89488, 89530, 89536, 89580, 89586, rs11564146_SNP 89495, 89500, 89543, 89548, 89594, 89599, 89502, 89504, 89550, 89552, 89601, 89609, 89510, 89519, 89558, 89566, 89613, 89618, 89522, 89527, 89568, 89570, 89620, 89622, 89630-89667 89668-89707 89708-89749 12:40360610_A_G 89750-89784 89785-89812 89813-89846 rs10784536_REF 12:40360610_A_G 89757-89760, 89786, 89789- 89816, 89819, rs10784536_SNP 89763, 89765, 89793, 89797- 89821-89822, 89772, 89774, 89798, 89802, 89828-89829, 89776, 89778, 89804, 89883- 89832, 89834, 89847-89882 89914 89836, 89842, 89915-89954 12:40360877_CTGCCTAT_C 89955-90001 90002-90051 90052-90103 rs11277474_REF 12:40360877_CTGCCTAT_C 89957, 89971, 90004, 90019, 90055, 90070, rs11277474_SNP 89974, 89984, 90022, 90038, 90074, 90090, 89997, 90104- 90047, 90144- 90101, 90186- 90143 90185 90229 12:40360895_G_A 89967-89970, 90009, 90012, 90053, 90060, rs7971919_REF 89981-89982, 90015-90018, 90063, 90066- 89986-89987, 90030-90031, 90069, 90082- 89994-89995, 90035-90037, 90083, 90087- 89998, 90108- 90044-90045, 90089, 90092, 90109, 90114, 90048, 90148- 90096-90097, 90125, 90127, 90149, 90155, 90100, 90190- 90230-90260 90166, 90168, 90191, 90197, 90261-90291 90208, 90220, 90292-90322 12:40360895_G_A 89967-89969, 90012, 90015- 90063, 90066, rs7971919_SNP 89981, 89994, 90016, 90030, 90082, 90092, 90232, 90245, 90044, 90261, 90096, 90292, 90248, 90250, 90264, 90276, 90294, 90306, 90257, 90323- 90279, 90288, 90309, 90315, 90360 90361-90400 90401-90442 12:40360980_T_G 90443-90449 90450-90453 90454-90460 rs7962116_REF 12:40360980_T_G 90443-90449, 90450-90453, 90454-90460, rs7962116_SNP 90461-90474 90475-90477 90478-90486 12:40361242_TTACA_T 90487-90488 rs67512688_REF 12:40361242_TTACA_T 90487-90488 rs67512688_SNP 12:40361614_G_A 90489-90517 90518-90530 90531-90550 rs2896978_REF 12:40361614_G_A 90498, 90516- 90535, 90558- rs2896978_SNP 90517, 90551- 90559 90557 12:40361674_A_T 90560-90607 90608-90657 90658-90709 rs2404836_REF 12:40361674_A_T 90564, 90569- 90612, 90614, 90662, 90664, rs2404836_SNP 90570, 90574, 90616, 90619- 90666, 90669, 90580, 90586, 90620, 90624, 90673, 90675, 90588, 90593, 90636, 90638, 90685, 90688, 90597, 90600, 90643, 90647, 90690, 90695, 90710-90747 90748-90787 90788-90829 12:40362405_T_C 90830-90876 90877-90926 90927-90978 rs11176195_REF 12:40362405_T_C 90840, 90842, 90888, 90890, 90931, 90937, rs11176195_SNP 90856, 90858- 90894, 90907- 90940, 90942, 90860, 90866, 90910, 90916, 90946, 90960- 90869, 90871, 90921, 90925, 90962, 90968, 90875, 90979- 91017-91056 90973, 91057- 91016 91098 12:40362446_A_G 91099-91146 91147-91192 91193-91242 rs4767973_REF 12:40362446_A_G 91103, 91108, 91151, 91156, 91197, 91207- rs4767973_SNP 91114, 91118, 91161-91162, 91208, 91212, 91120-91122, 91166, 91169- 91216, 91227, 91130, 91139, 91170, 91181, 91234-91235, 91145, 91243- 91186, 91191, 91239, 91241, 91280 91281-91320 91321-91362 12:40362519_C_T 91363-91404 91405-91438 91439-91477 rs17444202_REF 12:40362519_C_T 91369, 91384, 91411, 91425, 91450, 91461, rs17444202_SNP 91387, 91393, 91434, 91501- 91467, 91473, 91400, 91478- 91507 91508-91522 91500 12:40362670_C_A 91523-91570 91571-91619 91620-91671 rs4768236_REF 12:40362670_C_A 91535, 91540, 91583, 91588, 91633, 91638, rs4768236_SNP 91543, 91546, 91591, 91594, 91641, 91644, 91558-91559, 91603, 91608- 91653, 91657, 91562, 91566, 91609, 91612, 91659-91660, 91569-91570, 91619, 91710- 91663, 91671, 91672-91709 91749 91750-91791 12:40362885_T_C 91792-91814 91815-91825 91826-91843 rs4768237_REF 12:40362885_T_C 91795-91796, 91815-91816, 91827-91828, rs4768237_SNP 91799, 91801, 91818-91820, 91831, 91835, 91803-91804, 91822-91825, 91837-91839, 91808-91809, 91866-91883 91841-91843, 91811, 91813, 91884-91905 91844-91865 12:40363312_A_ACATG 91906-91907 rs58724345_SNP 12:40363384_AT_A 91908-91913 rs397897325_REF 12:40363384_AT_A 91910, 91913- rs397897325_SNP 91914 12:40363528_A_G 26687, 26689, 26729, 26731, 26770, 26772, rs33962975_REF 26694, 26704, 26733, 26738, 26774, 26776, 26720, 91915- 26747, 91954- 26781, 91996- 91953 91995 92039 12:40363528_A_G 91916, 91918- 91955, 91957, 91996, 91998, rs33962975_SNP 91919, 91921, 91959, 91964, 92002, 92006, 91923, 91926, 91976, 91985, 92018, 92029, 91938, 91946, 91994, 92078- 92038, 92118- 92040-92077 92117 92159 12:40363731_A_G 92160-92179 92180-92185 92186-92195 rs7134408_REF 12:40363731_A_G 92160, 92163, 92185, 92234- 92186, 92259- rs7134408_SNP 92170, 92172, 92258 92292 92177, 92196- 92233 12:40364243_T_G 92293-92339 92340-92381 92382-92429 rs11835105_REF 12:40364243_T_G 92296, 92298- 92341, 92343- 92384, 92388, rs11835105_SNP 92300, 92304, 92345, 92353- 92396-92398, 92306, 92309- 92355, 92370- 92404, 92408, 92311, 92328, 92371, 92379, 92417-92418, 92430-92467 92468-92507 92426, 92508- 92549 12:40364289_T_TA 92550-92597 92598-92647 92648-92699 rs137858615_REF 12:40364289_T_TA 92550-92551, 92598, 92602, 92648, 92652, rs137858615_SNP 92554, 92556, 92604, 92608, 92654, 92659, 92571, 92573- 92619, 92621, 92672, 92680, 92574, 92577, 92625, 92629, 92685, 92689, 92589, 92700- 92638, 92738- 92691, 92778- 92737 92777 92819 12:40364489_G_A 92820-92859 92860-92880 92881-92911 rs10878434_REF 12:40364489_G_A 92825, 92829, 92927-92930 92895, 92931- rs10878434_SNP 92838, 92840, 92938 92846, 92912- 92926 12:40364516_C_CT 92939-92942 92943 92944-92945 rs35403247_REF 12:40364850_T_C 26835, 26846, 26978, 26980, 26904, 26907, rs3761863_REF 26939, 26949, 26998, 92980- 27019, 27037, 26959, 92946- 93009 27039, 93010- 92979 93044 12:40364850_T_C 26939, 26949, 26978, 26980, 27019, 27037, rs3761863_SNP 26959, 92952, 26998, 92986, 27039, 93011, 92958, 92972, 92989, 93006, 93017, 93033, 92976, 93045- 93082 93083-93122 93123-93164 12:40365793_G_A 93165-93210 93211-93253 93254-93300 rs17444223_REF 12:40365793_G_A 93180, 93185, 93222, 93237- 93260, 93266, rs17444223_SNP 93187, 93192- 93238, 93241, 93283, 93286, 93193, 93196, 93249, 93330- 93296, 93343- 93201, 93206, 93342 93367 93209, 93301- 93329 12:40366046_T_A 93368-93409 93410-93443 93444-93483 rs12426362_REF 12:40366046_T_A 93371, 93377, 93412, 93415, 93447, 93453, rs12426362_SNP 93391, 93399, 93427, 93521- 93474, 93552- 93406, 93484- 93551 93588 93520 12:40366178_A_G 93589-93635 93636-93685 93686-93737 rs17466605_REF 12:40366178_A_G 93591-93592, 93638, 93648, 93688, 93695, rs17466605_SNP 93601, 93605- 93652-93653, 93699-93700, 93606, 93621- 93666, 93669- 93704-93705, 93622, 93625, 93670, 93673, 93718, 93721, 93633, 93738- 93682-93683, 93725, 93734, 93775 93776-93815 93816-93857 12:40367205_A_G 93858-93867 , 93868-93869 rs3789328_REF 12:40367205_A_G 93858-93859, 93897-93907 93908-93924 rs3789328_SNP 93863-93864, 93867, 93870- 93896 12:40367513_A_T 93925-93934 93935-93938 93939-93944 rs12426498_REF 12:40367513_A_T 93925, 93945- 93954-93957 93958-93963 rs12426498_SNP 93953 12:40367556_A_G 93964-93977 93978-93987 93988-94001 rs4768238_REF 12:40367556_A_G 93965, 93967, 93979, 93983, 93989, 93991, rs4768238_SNP 93972, 93974, 93985, 93987, 93997, 93999, 93977, 94002- 94027-94037 94001, 94038- 94026 94052 12:40367861_T_C 28203, 94053- 94090-94122 28263, 28266, rs66737902_REF 94089 94123-94164 12:40367861_T_C 94059, 94063, 94096, 94098, 94130, 94140, rs66737902_SNP 94067, 94070- 94102, 94105- 94143, 94146, 94071, 94080, 94106, 94109, 94148, 94152, 94082-94083, 94115-94116, 94155, 94157, 94086, 94165- 94119, 94201- 94160-94161, 94200 94236 94237-94278 12:40368129_A_C 94279-94311 94312-94345 94346-94381 rs10878441_REF 12:40368129_A_C 94282, 94288, 94312, 94316, 94346-94347, rs10878441_SNP 94292, 94304, 94322, 94327, 94351, 94357, 94382-94416 94340, 94417- 94375, 94450- 94449 94484 12:40368149_C_T 28531, 28536, 28556, 28560, 28577, 28580, rs3886747_REF 28539, 94282, 94312, 94316, 28585, 28587, 94288, 94292, 94322, 94327, 94346-94347, 94295, 94297, 94330, 94333, 94351, 94357, 94302, 94304, 94337, 94340, 94362, 94365, 94310, 94485- 94342, 94344, 94368, 94370, 94517 94518-94547 94372, 94375, 94377, 94380, 94548-94581 12:40368149_C_T 94295, 94297, 94330, 94333, 94365, 94368, rs3886747_SNP 94302, 94310, 94337, 94342, 94370, 94377, 94488, 94490, 94344, 94522, 94380, 94551, 94498-94499, 94530-94531, 94556, 94561- 94510, 94582- 94540-94541, 94562, 94573, 94603 94604-94616 94617-94636 12:40368501_G_C 28607-28608, 28616, 28663, 28621-28622, rs1365770_REF 28630, 28634, 28666, 28672, 28693, 28697, 28641, 28644, 28675, 28683, 28704, 28715, 28654, 94637- 94665-94685 28721, 94686- 94664 94709 12:40368501_G_C 94637-94638, 94665-94667, 94686-94688, rs1365770_SNP 94640, 94643, 94671, 94685, 94694-94695, 94646, 94652, 94738-94759 94760-94785 94658, 94664, 94710-94737 12:40368690_CT_C 28783, 94786- 94798-94807 94808-94819 rs367772598_REF 94797 12:40368690_C_CTTT 28783, 94786- 94798-94807 94808-94819 rs386376255_REF 94797 12:40368744_T_C 28925, 28945, 94850-94877 94878-94911 rs10784548_REF 94820-94849 12:40368744_T_C 94825, 94829, 94851, 94855- 94879, 94884- rs10784548_SNP 94835, 94839- 94856, 94860, 94885, 94889- 94840, 94846, 94865, 94868- 94891, 94900, 94912-94944 94869, 94945- 94976-95010 94975 12:40369805_C_T 95011-95043 95044-95050 95051-95066 rs1465527_REF 12:40369805_C_T 95018, 95021, 95045, 95050 95055, 95057, rs1465527_SNP 95023, 95033, 95063 95037 12:40369814_C_T 95011-95014, 95046-95049, 95051-95052, rs1465528_REF 95016-95017, 95078-95082 95054, 95056, 95019-95020, 95058-95061, 95022, 95024- 95065-95066, 95025, 95027- 95083-95090 95028, 95030- 95031, 95035- 95036, 95038- 95039, 95041- 95043, 95067- 95077 12:40369814_C_T 95019, 95028, 95078-95080, 95084-95085, rs1465528_SNP 95039, 95070- 95082 95087, 95089- 95072, 95074, 95091 95076 12:40369850_G_T 95092-95121 95122-95136 95137-95158 rs17444285_REF 12:40369850_G_T 95096, 95101, 95124, 95127, 95142, 95149, rs17444285_SNP 95106-95107, 95131, 95133- 95152-95153, 95110, 95112, 95136 95155-95158, 95116-95117, 95162 95120-95121, 95159-95161 12:40370171_C_G 95163-95210 95211-95260 95261-95312 rs7968048_REF 12:40370171_C_G 95165, 95169- 95213, 95217, 95267, 95269, rs7968048_SNP 95171, 95180, 95219, 95228, 95277, 95279, 95182, 95191, 95230, 95239, 95281, 95294- 95195, 95206, 95243-95244, 95295, 95306- 95208, 95313- 95255-95256, 95307, 95312, 95350 95351-95390 95391-95432 12:40370967_T_G 95433-95443 , 95444 rs11609433_REF 12:40370967_T_G 95433, 95438, 95458-95468 95444, 95469- rs11609433_SNP 95445-95457 95483 12:40371783_GAAAAAAAAA 95484-95491 95492-95499 95500-95509 G rs776238897_REF 12:40371783_GAAAA_G 95484-95491 95492-95499 95500-95509 rs149833872_REF 12:40371783_GA_G 95484-95491 95492-95499 95500-95509 rs1299545494_REF 12:40371801_A_C 95510-95514 95515-95516 95517-95519 rs4374003_REF 12:40371801_A_C 95510-95514, 95515-95516, 95517-95519, rs4374003_SNP 95520-95522 95523-95525 95526-95530 12:40372127_A_G 95531-95559 95560-95573 95574-95596 rs7962370_REF 12:40372127_A_G 95538, 95543, 95563-95564, 95578-95579, rs7962370_SNP 95548, 95551- 95570-95572, 95584, 95589, 95552, 95554- 95635-95666 95591, 95594, 95555, 95597- 95667-95704 95634 12:40224997-40225496 14, 21, 26, 31, 67, 79, 84, 96, 117, 129, 134, Up to 500 bp Downstream 46, 151, 153, 244, 357-358, 147-148, 268, to Exon 51 168, 173, 193, 361-363, 367, 293, 295-296, 310, 313-314, 370, 375, 381, 397, 408-409, 318, 320-321, 383, 499, 508, 412, 414, 418, 326, 333, 343, 512, 517, 520, 420, 422, 433 451, 454, 460, 529-530, 535 435, 558, 562- 464, 469, 478, 540, 542, 657 563, 568 571, 480-481, 486, 659, 663, 666- 580, 586, 590, 491, 609, 611, 667, 670, 681, 592, 594, 707- 615, 618-619, 683, 685, 690, 708, 710, 717- 622, 630, 633, 96490-97286 718, 721, 732, 636, 642, 734, 740, 742, 95705-96489 97287-98083 12:40225283-40225554 95705, 95707- 96490, 96492- 97287, 97289- Intron 1 95709, 95711, 96494, 96496, 97291, 97293, 95713-95714, 96498-96499, 97295-97296, 95716-95720, 96501-96505, 97298-97302, 95722-95725, 96507-96510, 97304-97307, 95730, 95733, 96515, 96518, 97312, 97315, 95735, 95738, 96520, 96523, 97317, 97320, 95740, 95742- 96525, 96527- 97322, 97324- 95743, 95746- 96528, 96531- 97325, 97328- 95753, 95757, 96538, 96542, 97335, 97339, 95759-95763, 96544-96548, 97341-97345, 95771, 95776- 96556, 96562- 97353, 97358- 95777, 95781- 96563, 96566- 97359, 97362- 95782, 95785, 96567, 96570, 97363, 97366, 95792, 95795- 96577, 96580, 97373, 97376, 95798, 95800, 96582-96584, 97378-97380, 95802-95803, 96586, 96588- 97382, 97384- 95805, 95810- 96589, 96591, 97385, 97387, 95811, 95815, 96594, 96597- 97392-97393, 95824, 95826, 96598, 96602, 97397, 97406, 95831-95833, 96611, 96614, 97409, 97414- 95835-95838, 96619-96621, 97416, 97418- 95842, 95844- 96623-96626, 97421, 97426, 95845, 95847- 96630, 96632- 97428-97429, 95849, 95852, 96633, 96635- 97431-97433, 95854-95855, 96637, 96640, 97436, 97438- 95860, 95863- 96642-96643, 97439, 97445, 95864, 95867, 96648, 96651- 97448-97449, 95873, 95875, 96652, 96655, 97453, 97459, 95878, 95880- 96661, 96663, 97461, 97464, 95882, 95885- 96666, 96668- 97466-97468, 95891, 95893- 96670, 96673- 97471-97477, 95894, 95897, 96679, 96681- 97479-97480, 95906, 95909, 96682, 96685, 97483, 97492, 95916, 95918, 96694, 96697, 97495, 97505, 95920, 95924, 96707, 96709, 97507, 97509, 95927, 95933, 96711, 96715, 97513, 97516, 95943, 95947- 96718, 96724, 97522, 97532, 95948, 95950, 96734, 96738- 97536-97538, 95952, 95954, 96740, 96742- 97540-97542, 95956, 95958, 96744, 96746, 97544, 97547- 95961-95962, 96749-96750, 97548, 97550, 95964, 95967, 96752, 96755- 97554, 97556, 95970, 95972, 96756, 96758, 97558, 97561- 95975-95977, 96761-96763, 97563, 97566, 95979, 95983, 96766, 96770, 97570, 97574- 95987-95990, 96774-96777, 97577, 97579- 95992-95994, 96779-96781, 97581, 97586, 95999, 96009, 96786, 96797, 97596, 97600- 96013-96014, 96801-96802, 97601, 97603, 96017, 96024, 96804, 96814, 97613, 97617, 96027, 96030, 96818, 96820, 97619, 97622- 96032-96033, 96823-96824, 97623, 97625, 96035, 96037- 96826, 96828- 97627-97629, 96038, 96042, 96830, 96834, 97633, 97635- 96044-96045, 96836-96837, 97636, 97641, 96050, 96054- 96842, 96846- 97644, 97646, 96055, 96058, 96847, 96851, 97650, 97659- 96068-96072, 96859, 96861- 97663, 97665, 96074, 96078- 96865, 96867, 97668-97675, 96085, 96087, 96871-96878, 97677, 97679, 96089, 96091- 96880, 96882, 97681-97683, 96093, 96099, 96884-96887, 97689, 97693, 96102, 96104- 96892, 96896, 97695-97696, 96105, 96109- 96898-96899, 97700-97701, 96110, 96114- 96903-96904, 97705-97708, 96117, 96119- 96908-96912, 97710-97711, 96120, 96125, 96914-96915, 97718, 97720, 96127, 96130, 96921, 96923, 97723, 97725, 96132, 96134, 96926, 96928, 97727, 97730, 96137, 96141- 96930, 96933, 97734-97737, 96145, 96148, 96937-96940, 97741, 97743, 96151, 96153, 96944, 96946, 97745, 97752, 96160, 96162, 96948, 96955, 97754, 97765, 96173, 96178, 96957, 96968, 97770, 97773, 96183, 96186, 96972, 96975, 97776, 97780, 96188, 96196- 96978, 96982, 97787-97788, 96197, 96200, 96989-96990, 97791, 97794- 96203-96204, 96993, 96995- 97795, 97800- 96209-96212, 96996, 97001- 97803, 97805- 96214-96215, 97004, 97006- 97806, 97809, 96218, 96220- 97007, 97010, 97811-97812, 96221, 96225, 97012-97013, 97816, 97823, 96234, 96237- 97017, 97023, 97825-97826, 96238, 96240, 97025-97027, 97828, 97830- 96242-96243, 97029-97030, 97831, 97834- 96247-96248, 97033-97035, 97836, 97838, 96250, 96252, 97037, 97039, 97841, 97844, 96255, 96258, 97042, 97044, 97846, 97848- 96260, 96262- 97046-97047, 97849, 97857- 96263, 96271- 97055-97057, 97858, 97861- 96273, 96276- 97060-97061, 97862, 97867- 96277, 96283- 97067-97068, 97868, 97870- 96284, 96286- 97070-97071, 97871, 97873, 96287, 96289, 97073, 97078, 97878, 97890- 96304-96305, 97091-97092, 97891, 97893- 96307-96311, 97094-97098, 97897, 97903, 96316, 96318, 97104, 97106, 97905, 97907, 96320-96321, 97108, 97110, 97909, 97911- 96323-96325, 97112-97114, 97913, 97915, 96327, 96329, 97116, 97118, 97917, 97922- 96333-96334, 97122-97123, 97923, 97931- 96342-96343, 97131-97132, 97932, 97934, 96345, 96348- 97134, 97138, 97938, 97940- 96350, 96353, 97140-97141, 97941, 97944, 96355-96357, 97144, 97147- 97947-97949, 96359, 96363- 97149, 97151, 97951, 97956- 96365, 96367, 97156-97158, 97958, 97960, 96370-96371, 97160, 97163- 97963-97964, 96373-96374, 97164, 97166- 97966-97967, 96379, 96381- 97167, 97172, 97972, 97974, 96383, 96385, 97174, 97176- 97976-97977, 96390-96392, 97177, 97179, 97979, 97983- 96395, 96397- 97183-97185, 97985, 97988, 96398, 96403, 97188, 97191- 97991-97992, 96413, 96417, 97192, 97197- 97996-97997, 96420, 96425- 97198, 97208, 98006, 98010, 96427, 96429- 97212, 97215, 98013, 98018- 96430, 96432, 97219-97221, 98019, 98021- 96434, 96436, 97223-97224, 98022, 98024, 96441, 96443, 97226, 97228, 98026, 98029, 96446-96448, 97231, 97236, 98034, 98036, 96450-96452, 97238, 97242- 98040-98045, 96454-96457, 97247, 97249- 98047-98050, 96460-96461, 97252, 97255, 98053, 98055, 96463, 96465, 97257, 97259, 98057, 98059, 96467-96476, 97261, 97263, 98061, 98064- 96478-96480, 97266-97273, 98071, 98073- 96483-96489, 97275-97277, 98075, 98078- 98084-98199 97280-97286, 98083, 98316- 98200-98315 98431 12:40366506-40367005 98432-98857 98858-99333 99334-99715 Intron 49 up to 500 bp Upstream to Exon 50 12:40367078-40367643 93964-93967, 93978-93987, 93988-93991, Intron 50 93969-93977, 99872-100053 93993-93998, 99716-99871 100054-100193 12:40225641-40226140 100194-100907 100908-101671 101672-102343 Intron 2 up to 500 bp Downstream to Exon 2 12:40235126-40235625 102344-102742 102743-103177 103178-103604 Intron 3 up to 500 bp Upstream to Exon 4 The indicated locations listed in column 1 of the Table 1 are based on gnomAD v3 database and UCSC Genome Browser assembly ID: hg38, Sequencing/Assembly provider ID: Genome Reference Consortium Human GRCh38.p12 (GCA_000001405.27). Assembly date: December 2013 initial release; December 2017 patch release 12. The SNP details are indicated by the listed SNP ID Nos. (“rs numbers”), which are based on the NCBI 2018 database of Single Nucleotide Polymorphisms (dbSNP)). The indicated DNA mutations are associated with Transcript Consequence NM_198578 as obtained from NCBI RefSeq genes.

Examples are provided below to facilitate a more complete understanding of the invention. The following examples illustrate the exemplary modes of making and practicing the invention. However, the scope of the invention is not limited to specific embodiments disclosed in these Examples, which are for purposes of illustration only.

Experimental Details Example 1: LRRK2 On-Target Activity Anaylsis

Guide sequence portions comprising 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 are screened for high on target activity using SpCas9 in HeLa cells. On target activity is determined by DNA capillary electrophoresis analysis.

Example 2: LRRK2 Editing Strategy Analysis Using SpCas9

Point mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most frequent cause of familial, autosomal dominant Parkinson's disease. In order to develop an editing strategy requiring only one guide RNA molecule to target and inactivate an LRRK2 mutant allele, guide RNA molecules having guide sequence portions which target SNPs residing in the coding region of the gene were screened. Specifically, guides having one of nine (9) different guide sequence portions, each of which target a CRISPR complex to one of four different SNP positions (rs1878245, rs1427263, rs1878405, or rs3761863), were screened in A549 cells that express LRRK2 to determine if the guides could lead to the formation of indels and/or frameshift mutations in the LRRK2 coding sequence that could result in expression of a truncated protein or induce nonsense mediated decay (NMD).

Briefly, pre-assembled RNPs composed of 105 pmole SpCas9 CRISPR nuclease protein and 120 pmole of a 20-nucleotide sgRNA (Table 2) were mixed with 100 pmole of electroporation enhancer (IDT-1075916) and electroporated into 2×10⁵ A549 cells using the SF primary cell 4D-nucleofector X Kit S (PBC2-00675, Lonza) and applying the CM-130 program. A fraction of cells was harvested 72 hours post electroporation, genomic DNA was extracted, and amplicons generated from the extracted DNA was subjected to next-generation sequencing (NGS) analysis. As shown in the results presented in FIG. 1 , the tested guide molecules depicted high editing activity at the site of the targeted SNPs, ranging from 76%-97% on-target activity.

To assess the effect of the editing on the level of LRRK2 expression, total RNA was extracted from A549 cells seven (7) days post-electroporation and the level of LRRK2 mRNA was measured by qRT-PCR. A decrease in the level of LRRK2 mRNA was detected only for guide molecules “g8” and “g9_alt,” measuring 60% and 40% editing respectively. The editing pattern for the rest of the guide molecules indicates that truncated protein expression could be produced as the result of a frameshift mutation.

TABLE 2 Sequences of 20-nucleotide guide sequence portions that target the human LRRK2 coding sequence. Location SNP Guide Sequence PAM Exon 5 rs1878245 g1_alt UUCUUCAUCCAGUAUCAGCA AGG (T > C) (SEQ ID NO: 33653) g2_alt UAAAAUCACCUUGCUGAUAC UGG (SEQ ID NO: 33642) Exon 34 rs1427263 g3 CUACGCGAAAUAAUGCCCUU AGG (C > A) (SEQ ID NO: 61673) g4 UACGCGAAAUAAUGCCCUUA GGG (SEQ ID NO: 61681) g5 AUAAUGCCCUUAGGGUGUUU UGG (SEQ ID NO: 61661) Exon 43 rs1878405 g6 AAUUUCUCAACCAUAGGCCA UGG (G > A) (SEQ ID NO: 80365) g7 AUUUCUCAACCAUAGGCCAU GGG (SEQ ID NO: 80368) g8 UUUCUCAACCAUAGGCCAUG GGG (SEQ ID NO: 80384) Exon 49 rs3761863 g9_alt GGUAACGGUAAAAGAAAACA AGG (T > C) (SEQ ID NO: 93060)

Example 3: LRRK2 Editing Strategy Analysis Using Alternate CRISPR Nucleases

The editing activities of two alternate CRISPR nucleases, OMNI-159 (SEQ ID NO: 103612) and OMNI-79 V5570 (SEQ ID NO: 103613), were preliminarily assessed with select guides in HeLa cells (FIG. 3 and FIG. 4 ). These initial data suggest that the LRRK2-targeting guide sequence portions provided herein can be utilized with a variety of different CRISPR nucleases.

TABLE 3 Sequences of guide sequence portions that target the human LRRK2 coding sequence used with alternate CRISPR nucleases. OMNI CRISPR SNP Targeted Guide Name Guide Sequence Portion PAM Nuclease rs3761863(T > C) g11_alt UCUAGGGAGGUAACGGUAAAGG AAAACA OMNI-159 (SEQ ID NO: 93150) rs1427263(C > A) g5_alt AAAUAAUUCCCUUAGGGUGUUU TGGACA OMNI-159 (SEQ ID NO: 61854) rs10878245(T > C) g13_alt AUAUCACUUUCUUCAUCCAGUA TCAGCA OMNI-159 (SEQ ID NO: 33706) rs10878245(T > C) g1_alt CUUUCUUCAUCCAGUAUCAGCA AGGTG OMNI-79 V5570 (SEQ ID NO: 33719) rs1427263(C > A) g4_alt UCUACGCGAAAUAAUUCCCUUA GGGTG OMNI-79 V5570 (SEQ ID NO: 61889)

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1. A method for modifying in a cell a mutant allele of the Leucine-rich repeat serine/threonine-protein kinase 2 (LRRK2) gene having a mutation associated with monogenic Parkinson's disease, the method comprising introducing to the cell a composition comprising: at least one CRISPR nuclease or a sequence encoding a CRISPR nuclease; and a first RNA molecule comprising a guide sequence portion having 17-50 nucleotides or a nucleotide sequence encoding the same, wherein a complex of the CRISPR nuclease and the first RNA molecule affects a double strand break in the mutant allele of the LRRK2 gene.
 2. The method of claim 1, wherein the first RNA molecule targets the CRISPR nuclease to the mutation associated with monogenic Parkinson's disease, wherein the mutation associated with monogenic Parkinson's disease is any one of 12:40225041_C_T, 12:40225058_C_A, 12:40225088_G_A, 12:40225159_G_A, 12:40225233_G_C, 12:40225558_C_T, 12:40225618_T_C, 12:40225628_G_A, 12:40225631_T_C, 12:40232303_A_C, 12:40232345_G_C, 12:40232380_A_C, 12:40232385_T_C, 12:40235634_T_C, 12:40235646_C_G, 12:40235660_A_G, 12:40238078_A_G, 12:40240517_C_T, 12:40240531_T_C, 12:40240594_G_C, 12:40240602_T_C, 12:40243551_CAAT_C, 12:40243556_A_T, 12:40249822_T_G, 12:40249843_C_G, 12:40249854_C_T, 12:40249881_T_C, 12:40249908_A_G, 12:40251273_G_A, 12:40251303_G_A, 12:40251361_A_G, 12:40251366_C_A, 12:40251369_G_A, 12:40251495_C_T, 12:40252906_A_G, 12:40252929_G_A, 12:40252939_C_T, 12:40252952_T_G, 12:40252958_A_C, 12:40252968_G_A, 12:40252974_C_T, 12:40252978_C_T, 12:40252984_C_T, 12:40252988_T_C, 12:40253016_G_A, 12:40257311_A_G, 12:40257341_G_T, 12:40259507_A_G, 12:40259525_A_T, 12:40259534_G_T, 12:40259578_G_A, 12:40263806_A_G, 12:40263875_A_G, 12:40263900_G_A, 12:40263907_T_C, 12:40274646_CC_C, 12:40274881_T_C, 12:40274899_A_G, 12:40274917_G_C, 12:40277933_T_C, 12:40277968_A_C, 12:40277971_A_G, 12:40278025_T_A, 12:40278154_A_G, 12:40278167_C_T, 12:40283881_G_A, 12:40283915_T_C, 12:40283947_C_T, 12:40283966_G_A, 12:40283985_C_A, 12:40283990_T_C, 12:40284061_A_G, 12:40284076_C_A, 12:40284139_T_C, 12:40287363_C_A, 12:40287438_A_G, 12:40287461_A_G, 12:40293555_CTC_CCT, 12:40293607_C_T, 12:40293613_G_A, 12:40293624_G_C, 12:40293626_G_A, 12:40293644_A_G, 12:40294854_T_G, 12:40294862_T_C, 12:40294866_G_T, 12:40294878_C_T, 12:40295448_A_G, 12:40295463_A_G, 12:40295466_G_A, 12:40295535_A_T, 12:40295566_A_G, 12:40295574_A_C, 12:40295581_G_T, 12:40298330_A_G, 12:40298333_C_T, 12:40298346_G_A, 12:40298433_C_G, 12:40298479_G_T, 12:40298488_A_G, 12:40299125_A_G, 12:40299190_C_T, 12:40299196_A_C, 12:40299212_G_A, 12:40299255_T_C, 12:40302866_A_G, 12:40303952_C_A, 12:40303972_C_G, 12:40304000_G_A, 12:40304004_A_G, 12:40304035_A_C, 12:40304040_G_C, 12:40304040_G_T, 12:40304078_G_A, 12:40304141_C_T, 12:40305776_C_T, 12:40305781_T_A, 12:40305791_C_G, 12:40305871_G_A, 12:40305942_G_A, 12:40305946_T_A, 12:40308467_G_T, 12:40308476_A_G, 12:40308481_G_A, 12:40308543_GG_G, 12:40308632_C_A, 12:40308672_G_A, 12:40309174_G_A, 12:40309206_C_T, 12:40309225_A_C, 12:40309226_A_G, 12:40309230_A_G, 12:40309236_G_A, 12:40310434_C_A, 12:40310434_C_G, 12:40310434_C_T, 12:40310435_G_A, 12:40310436_C_T, 12:40310437_G_C, 12:40310440_T_G, 12:40310450_C_T, 12:40310461_G_A, 12:40310471_A_G, 12:40310476_GAT_G, 12:40310515_A_G, 12:40310530_C_T, 12:40310546_G_A, 12:40310554_G_A, 12:40310561_G_A, 12:40314043_G_A, 12:40314059_C_T, 12:40314062_A_G, 12:40314101_C_A, 12:40315266_T_A, 12:40319979_A_T, 12:40319998_T_C, 12:40320043_G_A, 12:40320054_G_C, 12:40320090_A_G, 12:40320094_A_G, 12:40320097_T_C, 12:40320119_A_G, 12:40321114_A_G, 12:40321123_T_C, 12:40322037_C_T, 12:40322038_G_A, 12:40322047_G_A, 12:40322047_G_T, 12:40322134_A_G, 12:40322137_A_C, 12:40322364_T_C, 12:40322386_G_T, 12:40322468_C_A, 12:40323244_C_G, 12:40323255_A_G, 12:40323256_T_C, 12:40323260_G_T, 12:40323270_G_T, 12:40323297_T_C, 12:40328416_G_C, 12:40335008_A_T, 12:40335031_G_A, 12:40335036_C_T, 12:40335037_G_A, 12:40335079_G_T, 12:40340349_C_G, 12:40340361_T_C, 12:40340380_T_C, 12:40340400_G_A, 12:40340404_T_C, 12:40340432_A_G, 12:40340436_A_T, 12:40346818_T_C, 12:40346826_ACTACT_A, 12:40346884_A_G, 12:40348402_G_GTGT, 12:40348475_A_G, 12:40348484_C_T, 12:40351537_A_G, 12:40351572_T_A, 12:40351579_C_T, 12:40351585_G_A, 12:40351605_G_C, 12:40351623_G_T, 12:40351667_C_A, 12:40351680_G_C, 12:40351720_G_C, 12:40351723_A_G, 12:40354304_T_C, 12:40354305_G_T, 12:40354410_G_A, 12:40356126_A_T, 12:40356145_C_T, 12:40359345_C_T, 12:40359422_A_G, 12:40363435_C_T, 12:40363440_C_T, 12:40363541_G_A, 12:40364843_G_A, 12:40364845_G_GGT, 12:40364884_G_A, 12:40364927_A_T, 12:40364960_A_G, 12:40365044_C_T, 12:40367012_T_A, 12:40367050_A_G, 12:40367648_AC_A, 12:40367656_G_T, 12:40367754_T_C, 12:40367849_G_A, 12:40367888_A_C, 12:40367988_C_T, 12:40368042_A_G, 12:40368073_A_C, 12:40368170_C_T, 12:40368262_C_T, 12:40368380_T_A, 12:40368486_G_T, 12:40368516_C_A, 12:40368534_T_G, 12:40368673_A_C, 12:40368736_C_A, 12:40368783_G_T, 12:40368800_T_C, 12:40368807_A_G, 12:40368834_A_G, 12:40368860_C_T, 12:40368905_A_T, 12:40368930_C_T, 12:40368947_C_T, 12:40368951_C_T, 12:40369044_T_C, 12:40369153_T_C, and 12:40369167_G_T; and/or wherein the guide sequence portion of the first RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 that targets a mutation associated with monogenic Parkinson's disease. 3-4. (canceled)
 5. The method of claim 1, wherein the first RNA molecule targets the CRISPR nuclease to a SNP position of the mutant allele, wherein the SNP position is any one of rs1388598, rs28365207, rs7294619, rs10878226, rs1491943, rs1491942, rs1491941, rs11175645, rs4567538, rs12230685, rs11175655, rs11175656, rs1402299410, rs780970090, rs1479481931, rs71078229, rs11175658, rs3057613, rs35642658, rs74324737, rs2708441, rs10631840, rs35991926, rs61931634, rs11175666, rs751672176, rs36024911, rs377568728, rs1279923177, rs2249281, rs10878244, rs138000936, rs10878245, rs10878246, rs10878247, rs954884, rs10878249, rs10784444, rs6581622, rs17519573, rs35987733, rs868476079, rs1399756142, rs1474763022, rs1450276224, rs1191097078, rs1425272990, rs1477924701, rs2404580, rs2404581, rs1388599, rs1388596, rs10878258, rs12371502, rs11175729, rs17443414, rs1486599371, rs7134379, rs7955902, rs1491939, rs1491938, rs7969677, rs2723265, rs7132171, rs10784451, rs10784452, rs1907633, rs1491936, rs1907632, rs2046928, rs2723264, rs17465870, rs10878278, rs11175766, rs11564183, rs11175769, rs17465912, rs17443552, rs17490817, rs11430638, rs4293189, rs10784461, rs7308720, rs10784462, rs10878284, rs11175780, rs11175784, rs11175787, rs144463374, rs73108329, rs12426891, rs4768224, rs11175793, rs10784470, rs12146857, rs11564207, rs7308193, rs10506150, rs76660489, rs10878299, rs373619252, rs10878302, rs11564129, rs7971935, rs11175836, rs4272849, rs10784485, rs10506151, rs7309197, rs7134572, rs11175843, rs147801391, rs376096297, rs7488873, rs11175845, rs7979341, rs1226327150, rs777452337, rs1339001348, rs11175847, rs112859457, rs17466080, rs11175852, rs10784486, rs73108368, rs398019201, rs17519846, rs17466150, rs11175862, rs4310676, rs11564150, rs4318033, rs4473003, rs1429478461, rs886049356, rs11564270, rs17519881, rs17466178, rs17466185, rs11175902, rs76033741, rs56325408, rs7957754, rs11175911, rs12369882, rs11175913, rs4483664, rs66487867, rs1491426750, rs5006481, rs139565575, rs150276517, rs72446556, rs1186032972, rs12820920, rs7966550, rs12314692, rs11175922, rs73102746, rs10784497, rs17443815, rs7957057, rs76607433, rs28365224, rs7962011, rs10878336, rs11175934, rs11175935, rs17491061, rs11175941, rs10878340, rs10784498, rs10878341, rs10784499, rs17484286, rs146071397, rs10878343, rs11564180, rs11175958, rs11175959, rs113772607, rs11564149, rs11175963, rs7305344, rs7133914, rs11175964, rs397805328, rs11175966, rs10784501, rs41286474, rs869212703, rs2069228, rs10784503, rs386376250, rs11564128, rs2896975, rs7302503, rs7137160, rs369794148, rs34709132, rs10467144, rs11175985, rs10784504, rs57421387, rs7973058, rs1427267, rs1427266, rs721713, rs721712, rs721711, rs721709, rs4768227, rs398098159, rs4767969, rs10878356, rs10784506, rs10878358, rs7132073, rs1427265, rs17466339, rs2114569, rs2114568, rs2162469, rs10784509, rs1896254, rs5797667, rs373676201, rs1896253, rs1896252, rs1427263, rs11176013, rs11564148, rs11564205, rs10748030, rs10748031, rs10748032, rs11564177, rs10748033, rs10735931, rs10748034, rs10878368, rs7307276, rs4768229, rs4768230, rs397968969, rs11564204, rs4423250, rs7137665, rs10878371, rs11176022, rs60750674, rs10878372, rs398055492, rs761931834, rs1415940047, rs17466430, rs11176030, rs3747891, rs7298930, rs17444089, rs10878377, rs12316801, rs17444096, rs11564175, rs2404832, rs17466486, rs11564267, rs17520244, rs10506152, rs11176052, rs11176053, rs17444124, rs11564203, rs7307562, rs17520251, rs11461395, rs10506153, rs73275761, rs963243, rs17444145, rs10878386, rs17466521, rs12367542, rs17520278, rs35031086, rs11834950, rs11829088, rs4465428, rs61007767, rs60776971, rs2404834, rs7302841, rs2404835, rs11356084, rs715403, rs715402, rs6581667, rs6581668, rs200521371, rs4767970, rs4767971, rs4767972, rs1427271, rs1427272, rs1427273, rs10735934, rs10732751, rs12306060, rs1365763, rs7963697, rs11316380, rs7963086, rs17491536, rs7956787, rs7956898, rs10506155, rs7954061, rs7957151, rs919174, rs3065921, rs919175, rs1035812, rs1365764, rs397724721, rs17461964, rs10784518, rs4768231, rs4768232, rs3943893, rs2162471, rs1035813, rs10459264, rs10459265, rs11564147, rs10715759, rs7296657, rs7312497, rs10715758, rs10784522, rs796827140, rs74467833, rs7131694, rs12311273, rs58911468, rs35847030, rs1365765, rs66513412, rs2896977, rs199606989, rs1365766, rs10878405, rs11176143, rs7294958, rs61915610, rs7294952, rs59096461, rs59980086, rs4768233, rs4768234, rs4768235, rs7314455, rs7313525, rs7313895, rs7314863, rs7303525, rs7132187, rs7307310, rs11176153, rs11176154, rs7311247, rs7137437, rs75439407, rs73107108, rs1307806388, rs10878409, rs10878410, rs138382676, rs11176161, rs10878412, rs10878413, rs183544696, rs189538849, rs11610569, rs142726158, rs1194726090, rs576444406, rs185723137, rs1179215763, rs71449759, rs61915613, rs140722234, rs191099092, rs113693842, rs112767992, rs34531686, rs12368227, rs12368229, rs3930031, rs7308626, rs890575, rs890575, rs10748040, rs61230125, rs11176165, rs7976715, rs7976724, rs7963987, rs2162472, rs767501990, rs552326490, rs1365381164, rs760911090, rs754028820, rs11564173, rs7306545, rs7306684, rs112259589, rs73277531, rs77689380, rs73277533, rs17462055, rs758880647, rs10784528, rs796949072, rs10784532, rs145580704, rs386376254, rs11836288, rs11564146, rs10784536, rs11277474, rs7971919, rs7962116, rs67512688, rs2896978, rs2404836, rs11176195, rs4767973, rs17444202, rs4768236, rs4768237, rs58724345, rs397897325, rs33962975, rs7134408, rs11835105, rs137858615, rs10878434, rs35403247, rs3761863, rs17444223, rs12426362, rs17466605, rs3789328, rs12426498, rs4768238, rs66737902, rs10878441, rs3886747, rs1365770, rs367772598, rs386376255, rs10784548, rs1465527, rs1465528, rs17444285, rs7968048, rs11609433, rs776238897, rs149833872, rs1299545494, rs4374003, and rs7962370; wherein the guide sequence portion of the first RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 that targets a SNP position of the mutant allele; wherein the SNP position is in an exon or intron of the LRRK2 mutant allele, or within 3,000 basepairs upstream of the transcription start site or within 3,000 basepairs downstream of the 3′ UTR of the LRRK2 mutant allele; and/or wherein the SNP position contains a heterozygous SNP. 6-9. (canceled)
 10. The method of claim 1, further comprising introducing to the cell a second RNA molecule comprising a guide sequence portion having 17-50 nucleotides or a nucleotide sequence encoding the same, wherein a complex of the second RNA molecule and a CRISPR nuclease affects a second double strand break in the LRRK2 gene.
 11. The method of claim 10, wherein the guide sequence portion of the second RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 other than the sequence of the first RNA molecule, wherein the second RNA molecule comprises a non-discriminatory guide portion that targets both functional and mutated LRRK2 alleles; wherein the second RNA molecule comprises a non-discriminatory guide portion that targets any one of Intron 1 of LRRK2, Intron 49 of LRRK2, Intron 2 of LRRK2, Intron 3 of LRRK2, a 3′ untranslated region (3′ UTR) of LRRK2, and an intergenic region downstream of LRRK2; wherein the second RNA molecule comprises a non-discriminatory guide portion that targets a sequence that is located within a genomic range selected from any one of 12:40224997-40225496, 12:40225283-40225554, 12:40366506-40367005, 12:40367078-40367643, 12:40225641-40226140, and 12:40235126-40235625; and/or wherein the second RNA molecule comprises a non-discriminatory guide portion that targets a sequence that is located up to 500 base pairs from an exon that is excised by the first and second RNA molecules. 12-15. (canceled)
 16. The method of claim 10, wherein an exon or a portion thereof is excised from the mutant allele of the LRRK2 gene.
 17. The method of claim 1, wherein the guide sequence portion of the first RNA molecule comprises 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion; or wherein the guide sequence portion comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606 modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion.
 18. (canceled)
 19. The method of claim 17, wherein the guide sequence portion provides higher targeting specificity to the complex of the CRISPR nuclease and the first RNA molecule relative to a guide sequence portion that has higher complementarity to the mutant allele of the LRRK2 gene.
 19. The method of claim 17, wherein the guide sequence portion provides higher targeting specificity to the complex of the CRISPR nuclease and the first RNA molecule relative to a guide sequence portion that has higher complementarity to the mutant allele of the LRRK2 gene.
 20. A modified cell obtained by the method of claim
 1. 21. The modified cell of claim 20, wherein the modified cell is a stem cell, a neuron, or a dopaminergic cell.
 22. A first RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606, or any one of SEQ ID NOs: 1-103,606 modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion.
 23. A composition comprising the first RNA molecule of claim 22 and at least one CRISPR nuclease.
 24. The composition of claim 23, further comprising a second RNA molecule comprising a guide sequence portion having 17-50 contiguous nucleotides, wherein the second RNA molecule targets a LRRK2 allele, and wherein the guide sequence portion of the second RNA molecule is a different sequence from the sequence of the guide sequence portion of the first RNA molecule.
 25. The composition of claim 24, wherein the guide sequence portion of the second RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs: 1-103,606, or any one of SEQ ID NOs: 1-103,606 modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully complementary target sequence of the guide sequence portion.
 26. A method for inactivating a mutant LRRK2 allele in a cell, the method comprising delivering to the cell the composition of claim
 23. 27. A method for treating monogenic Parkinson's disease, the method comprising delivering to a cell of a subject having monogenic Parkinson's disease the composition of claim
 23. 28. A method for inactivating a mutant LRRK2 allele in a cell, comprising delivering to the cell the composition of claim
 20. 29. A medicament comprising the composition of claim 23 for use in inactivating a mutant LRRK2 allele in a cell, wherein the medicament is administered by delivering to the cell the composition of claim
 23. 30. A method for treating ameliorating or preventing monogenic Parkinson's disease, comprising delivering to a cell of a subject having or at risk of having monogenic Parkinson's disease the composition of claim
 23. 31. A medicament comprising the composition of claim 23 for use in treating ameliorating or preventing monogenic Parkinson's disease, wherein the medicament is administered by delivering to a cell of a subject having or at risk of having monogenic Parkinson's disease the composition of claim
 23. 